Hemp cultivar named ‘05.09.24.S1’

ABSTRACT

The present disclosure provides a new and distinct hemp cultivar designated ‘05.09.24.S1’. The present disclosure relates to seeds of the hemp plant ‘05.09.24.S1,’ to plants and parts of the hemp plant ‘05.09.24.S1,’ and to methods for producing a hemp plant by crossing the hemp plant ‘05.09.24.S1’ with itself or other  cannabis  plants. The disclosure further relates to the morphological and physiological characteristics of the new and distinct hemp cultivar and its uses.

FIELD

The disclosure relates to a hemp varieties, hemp extracts,CBD-containing compositions, and methods of producing and using thesame.

BACKGROUND

Cannabis is a genus of flowering plants that includes at least threespecies, Cannabis sativa, Cannabis indica, and Cannabis ruderalis asdetermined by plant phenotypes and secondary metabolite profiles. Hemp,also known as industrial hemp, is a type of cannabis plant grownspecifically for the industrial uses of its derived products. In theUnited States, Cannabis is classified as hemp if it accumulates no morethan three-tenths of one percent (i.e., 0.3%) concentration oftetrahydrocannabinol (THC) at harvest maturity. Hemp plants can alsoaccumulate high levels of cannabidiol (CBD), which is used in a varietyof consumer goods, including food, drinks, dietary supplements andcosmetics.

Hemp production however, remains challenging for farmers. Feminizedseeds for high CBD producing lines with other agricultural traits arehighly desirable, but not yet widely available.

Thus, there remains a need for new hemp varieties to meet the growingdemand for fiber and CBD-based products.

BRIEF SUMMARY

This disclosure relates to a new and distinctive hemp cultivardesignated as ‘05.09.24.S1’. In some embodiments, the ‘05.09.24.S1’ isproduced by multiple rounds of self-fertilization of the parent ‘BIHEMP050924’ (USPP32,473). In other embodiments, the ‘05.09.24.S1’ is derivedfrom a Cannabis hybrid background.

The inventors reproduced the ‘05.09.24.S1’ cultivar through strategiccrosses and selections from proprietary lines. The ‘05.09.24.S1’ plantis maintained as a seed produced by self-pollination at the inventors'greenhouses, nurseries, fields and/or facilities in Salinas, Calif.

The present disclosure provides a new and distinctive hemp varietydesignated as ‘05.09.24.S1’. The present disclosure relates to the seedsof hemp variety ‘05.09.24.S1’, to the plants or parts of hemp variety‘05.09.24.S1’, to the plant cells of hemp variety ‘05.09.24.S1’, to theplants or plant parts or plant cells having all of the physiological andmorphological characteristics of hemp variety ‘05.09.24.S1’ and toplants or plant parts or plant cells having all of the physiological andmorphological characteristics of plant cells listed in Tables 1-5(and/or deposited under NCMA No. 202202006), including, but not limitedto, as determined at the 5% significance level when grown in the sameenvironmental conditions, including when grown side-by-side with acomparison or check cannabis and/or hemp plant.

The present disclosure relates to methods for producing a hemp plantand/or seed, by crossing the hemp variety ‘05.09.24.S1’ with itself oranother cannabis and/or hemp plant. A further aspect relates to hybridhemp plants, and hemp seeds produced by crossing the hemp variety‘05.09.24.S1’ with a cannabis and/or hemp plant.

Another aspect of the present disclosure is also directed to a method ofproducing a cannabinoid extract comprising contacting plants of the hempvariety ‘05.09.24.S1’ with a solvent or heat, and producing thecannabinoid extract.

In some embodiments, the present disclosure teaches a seed, plant, plantpart, or plant cell of hemp plant variety designated ‘05.09.24.S1’,wherein seed of the variety has been deposited under NCMA No. 202202006.In some embodiments, the present disclosure teaches that the plant partis an inflorescence and/or a flower.

In some embodiments, the present disclosure teaches a hemp plant or aplant part or a plant cell thereof, having all of the physiological andmorphological characteristics of the hemp plant variety designated‘05.09.24.S1’ listed in Tables 1-5 (and/or deposited under NCMA No.202202006) including, but not limited to, as determined at the 5%significance level when grown in the same environmental conditions,including when grown side-by-side with a comparison or check cannabisand/or hemp plant. In some embodiments, the present disclosure teaches ahemp plant, or a plant part or a plant cell thereof, having all of thephysiological and morphological characteristics of the hemp plant of thepresent disclosure.

In some embodiments, the present disclosure teaches a hemp plant, or aplant part or a plant cell thereof, having all of the physiological andmorphological characteristics of the hemp plant variety designated‘05.09.24.S1’, wherein said variety was deposited under NCMA No.202202006.

In some embodiments, the present disclosure teaches a tissue culture ofregenerable cells produced from the plant, plant part or plant cell ofthe present disclosure, wherein a new plant regenerated from the tissueculture has all of the morphological and physiological characteristicsof the hemp plant variety designated ‘05.09.24.S1’ listed in Tables 1-5(and/or deposited under NCMA No. 202202006) when grown under the sameenvironmental conditions. In some embodiments, the present disclosureteaches a hemp plant regenerated from the tissue culture of the presentdisclosure, said plant having all the morphological and physiologicalcharacteristics of the hemp of the present disclosure. In someembodiments, the present disclosure teaches a hemp plant regeneratedfrom the tissue culture, wherein the regenerated plant has all of thecharacteristics of the hemp plant variety designated ‘05.09.24.S1’,wherein seed of said variety was deposited under NCMA No. 202202006.

In some embodiments, the present disclosure teaches a method forproducing a hemp seed, comprising selfing the hemp plant of the presentdisclosure, and harvesting the resultant hemp seed. In some embodiments,the present disclosure teaches a hemp seed produced by the method of thepresent disclosure.

In some embodiments, the present disclosure teaches a method forproducing a hemp seed comprising crossing the hemp plant of the presentdisclosure with a second, distinct plant. In some embodiments, thepresent disclosure teaches an F₁ hemp seed produced by the method of thepresent disclosure. In some embodiments, the present disclosure teachesan F₁ hemp plant, or a part or a plant cell thereof, produced by growingthe seed of the present disclosure.

In some embodiments, the present disclosure teaches a method ofproducing a hemp plant derived from the variety ‘05.09.24.S1,’comprising: a) crossing the plant of the present disclosure, with itselfor a second plant to produce progeny seed; b) growing the progeny seedto produce a progeny plant and crossing the progeny plant with itself ora second plant to produce further progeny seed; and, optionally c)repeating step b) one or more times to produce the hemp plant derivedfrom the variety ‘05.09.24.S1’.

In some embodiments, the present disclosure teaches a method ofproducing a hemp plant derived from the variety ‘05.09.24.S1’, furthercomprising crossing the hemp plant derived from the variety‘05.09.24.S1,’ with a plant of a different genotype to produce seed of ahybrid plant derived from the hemp variety ‘05.09.24.S1’.

In some embodiments, the present disclosure teaches a method ofproducing a hemp plant derived from the variety ‘05.09.24.S1’,comprising propagating a vegetative cutting from a stock Cannabis plant,thereby producing the Cannabis plant derived from the hemp varietydesignated ‘05.09.24.S1’. In some embodiments, the stock Cannabis plantis a product of applying a plant breeding technique taught herein to‘05.09.24.S1’.

In some embodiments, the present disclosure teaches method for producinga Cannabis plant derived from a hemp variety designated ‘05.09.24.S1’,said method comprising: crossing a stock Cannabis plant with itself oranother Cannabis plant, thereby producing the Cannabis plant derivedfrom the hemp variety designated ‘05.09.24.S1’. In some embodiments, thestock Cannabis plant is a product of applying a plant breeding techniquetaught herein to ‘05.09.24.S1’.

In some embodiments, the present disclosure teaches a method forproducing nucleic acids, comprising isolating nucleic acids from theseed, plant, plant part, or plant cell of the present disclosure.

In some embodiments, the present disclosure teaches a hemp plantcomprising a single locus conversion and otherwise all of themorphological and physiological characteristics of the hemp plant‘05.09.24.S1’ when grown in the same environmental conditions. In someembodiments, the present disclosure teaches that the single locusconversion confers said plant with herbicide resistance. In someembodiments, the present disclosure teaches that the single locusconversion is an artificially mutated gene or nucleotide sequence. Insome embodiments, the present disclosure teaches that the single locusconversion is a gene that has been modified through the use of breedingtechniques taught in the present disclosure.

In some embodiments, the present disclosure teaches a cultivar of hempdesignated ‘05.09.24.S1’ as described and detailed in the presentdisclosure.

In some embodiments, the present disclosure teaches a method ofproducing a cannabinoid extract, said method comprising the steps (a)contacting the plant of the present disclosure with a solvent or heat,thereby producing a cannabinoid extract.

In some embodiments, the present disclosure teaches a dry, non-viableplant part, wherein seed of hemp plants producing said dry plant partshas been deposited under NCMA No. 202202006.

In some embodiments, the present disclosure teaches an assemblage ofdry, non-viable female inflorescences from a hemp plant varietydesignated ‘05.09.24.S1’ wherein seed the variety has been depositedunder NCMA No. 202202006. In some embodiments, the present disclosureteaches that a dry, non-viable plant part is an inflorescence and/or aflower.

In some embodiments, the present disclosure teaches a hemp plant of thepresent disclosure is asexually reproduced. In some embodiments, thepresent disclosure teaches a hemp plant of the present disclosure iscapable of producing an asexual clone of said hemp plant. In someembodiments, the present disclosure teaches that the asexual clone iscapable of producing said hemp plant taught in the present disclosure.

A further embodiment relates to a method for developing a hemp plant ina hemp plant breeding program, comprising applying plant breedingtechniques comprising crossing, recurrent selection, mutation breeding,wherein said mutation breeding selects for a mutation that isspontaneous or artificially induced, backcrossing, pedigree breeding,marker enhanced selection, haploid/double haploid production, ortransformation to the hemp plant of ‘05.09.24.S1’, or its parts, whereinapplication of said techniques results in development of a hemp plant.

A further embodiment relates to a method of introducing a mutation intothe genome of hemp plant ‘05.09.24.S1’, said method comprisingmutagenesis of the plant, or plant part thereof, of ‘05.09.24.S1’,wherein said mutagenesis is selected from the group consisting oftemperature, long-term seed storage, tissue culture conditions, ionizingradiation, chemical mutagens, and targeting induced local lesions ingenomes, and wherein the resulting plant comprises at least one genomemutation and producing plants therefrom.

A further embodiment relates to a method of editing the genome of hempplant ‘05.09.24.S1’, wherein said method is selected from the groupcomprising zinc finger nucleases, transcription activator-like effectornucleases (TALENs), engineered homing endonucleases/meganucleases, andthe clustered regularly interspaced short palindromic repeat(CRISPR)-associated protein9 (Cas9) system, and plants producedtherefrom.

DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

The accompanying photographs depict characteristics of the new‘05.09.24.S1’ plants as nearly true as possible reproductions. Theoverall appearance of the ‘05.09.24.S1’ plants in the photographs maydiffer slightly from the color values described in the detailedbotanical description of Tables 1-5.

FIG. 1 shows a pedigree of the ‘05.09.24.S1’ plant with its breedinghistory.

FIG. 2 shows a bird-eye view of the ‘05.09.24.S1’ plants withhomozygosity in a greenhouse setting. Two rows of the uniform‘05.09.24.S1’ plants are located between two solid white lines (in themiddle of FIG. 2).

FIG. 3 shows an overall view of the ‘05.09.24.S1’ plant in a vegetativestage.

FIG. 4 shows a close view of portions of leaves of the ‘05.09.24.S1’plant in a vegetative stage.

FIG. 5 shows a close view of individual leaves of the ‘05.09.24.S1’plant in a vegetative stage.

FIG. 6 shows a close view of upper part (including flowers) of the‘05.09.24.S1’ plant with main axis dominance close to floral maturity.

DETAILED DESCRIPTION OF THE INVENTION

All publications, patents and patent applications, including anydrawings and appendices, are herein incorporated by reference to thesame extent as if each individual publication or patent application wasspecifically and individually indicated to be incorporated by reference.

The following description includes information that may be useful inunderstanding the present disclosure. It is not an admission that any ofthe information provided herein is prior art or relevant to thepresently claimed disclosures, or that any publication specifically orimplicitly referenced is prior art.

Definitions

As used herein, the verb “comprise” is used in this description and inthe claims and its conjugations are used in its non-limiting sense tomean that items following the word are included, but items notspecifically mentioned are not excluded.

As used herein, the term “about” refers to plus or minus 10% of thereferenced number, unless otherwise stated or otherwise evident by thecontext (such as when a range would exceed 100% of a possible value orfall below 0% of a possible value). For example, reference to anabsolute content of a particular cannabinoid of “about 1%” means thatthat cannabinoid can be present at any amount ranging from 0.9% to 1.1%content by weight.

The disclosure provides cannabis hemp plants. As used herein, the term“plant” refers to plants in the genus of Cannabis and plants derivedthereof. Such as cannabis plants produced via asexual reproduction,tissue culture, and via seed production.

The disclosure provides plant parts. As used herein, the term “plantpart” refers to any part of a plant including but not limited to theembryo, shoot, root, stem, seed, stipule, leaf, petal, flower,inflorescence, bud, ovule, bract, trichome, branch, petiole, internode,bark, pubescence, tiller, rhizome, frond, blade, ovule, pollen, stamen,and the like. The two main parts of plants grown in some sort of media,such as soil or vermiculite, are often referred to as the “above-ground”part, also often referred to as the “shoots”, and the “below-ground”part, also often referred to as the “roots”. Plant parts may alsoinclude certain extracts such as kief or hash, which include cannabisplant trichomes or glands. In some embodiments, plant part should alsobe interpreted as referring to individual cells from the plant.

As used herein, the term “plant cell” refers to any plant cell from acannabis plant. Plant cells of the present disclosure include cells froma cannabis plant shoot, root, stem, seed, stipule, leaf, petal,inflorescence, bud, ovule, bract, trichome, petiole, internode. In someembodiments, the disclosed plant cell is from a cannabis trichome.

The term “a” or “an” refers to one or more of that entity; for example,“a gene” refers to one or more genes or at least one gene. As such, theterms “a” (or “an”), “one or more” and “at least one” are usedinterchangeably herein. In addition, reference to “an element” by theindefinite article “a” or “an” does not exclude the possibility thatmore than one of the elements is present, unless the context clearlyrequires that there is one and only one of the elements.

The International Code of Zoological Nomenclature defines rank, in thenomenclatural sense, as the level, for nomenclatural purposes, of ataxon in a taxonomic hierarchy (e.g., all families are for nomenclaturalpurposes at the same rank, which lies between superfamily andsubfamily). While somewhat arbitrary, there are seven main ranks definedby the international nomenclature codes: kingdom, phylum/division,class, order, family, genus, and species. Further taxonomic hierarchiesused in this disclosure are described below.

The disclosure provides plant cultivars. As used herein, the term“cultivar” means a group of similar plants that by structural featuresand performance (i.e., morphological and physiological characteristics)can be identified from other cultivars within the same species.Furthermore, the term “cultivar” variously refers to a variety, strainor race of plant that has been produced by horticultural or agronomictechniques and is not normally found in wild populations. The termscultivar, variety, strain and race are often used interchangeably byplant breeders, agronomists and farmers.

The term “variety” as used herein has identical meaning to thecorresponding definition in the International Convention for theProtection of New Varieties of Plants (UPOV treaty), of Dec. 2, 1961, asRevised at Geneva on Nov. 10, 1972, on Oct. 23, 1978, and on Mar. 19,1991. Thus, “variety” means a plant grouping within a single botanicaltaxon of the lowest known rank, which grouping, irrespective of whetherthe conditions for the grant of a breeder's right are fully met, can bei) defined by the expression of the characteristics resulting from agiven genotype or combination of genotypes, ii) distinguished from anyother plant grouping by the expression of at least one of the saidcharacteristics and iii) considered as a unit with regard to itssuitability for being propagated unchanged.

The disclosure provides methods for obtaining plant lines. As usedherein, the term “line” is used broadly to include, but is not limitedto, a group of plants vegetatively propagated from a single parentplant, via tissue culture techniques, or a group of inbred or hybridplants which are genetically very similar due to descent from a commonparent(s) (e.g., by selfing of a genetically stable cultivar. A plant issaid to “belong” to a particular line if it (a) is a primarytransformant (T0) plant regenerated from material of that line; (b) hasa pedigree comprised of a T0 plant of that line; or (c) is geneticallyvery similar due to common ancestry (e.g., via inbreeding or selfing).In this context, the term “pedigree” denotes the lineage of a plant,e.g. in terms of the sexual crosses affected such that a gene or acombination of genes, in heterozygous (hemizygous) or homozygouscondition, imparts a desired trait to the plant.

As used herein, the term “inbreeding” refers to the production ofoffspring via the mating between relatives. The plants resulting fromthe inbreeding process are referred to herein as “inbred plants” or“inbreds.”

The term single allele converted plant as used herein refers to thoseplants that are developed by a plant breeding technique calledbackcrossing wherein essentially all of the desired morphological andphysiological characteristics of an inbred are recovered in addition tothe single allele transferred into the inbred via the backcrossingtechnique.

The disclosure provides samples. As used herein, the term “sample”includes a sample from a plant, a plant part, a plant cell, or from atransmission vector, or a soil, water or air sample.

The disclosure provides offspring. As used herein, the term “offspring”refers to any plant resulting as progeny from a vegetative or sexualreproduction from one or more parent plants or descendants thereof. Forinstance, an offspring plant may be obtained by cloning or selfing of aparent plant or by crossing two parent plants and include selfings aswell as the F1 or F2 or still further generations. An F1 is afirst-generation offspring produced from parents at least one of whichis used for the first time as donor of a trait, while offspring ofsecond generation (F2) or subsequent generations (F3, F4, etc.) arespecimens produced from selfings of F1's, F2's etc. An F1 may thus be(and usually is) a hybrid resulting from a cross between two truebreeding parents (true-breeding is homozygous for a trait), while an F2may be (and usually is) an offspring resulting from self-pollination ofsaid F1 hybrids.

The disclosure provides methods for crossing a first plant with a secondplant. As used herein, the term “cross”, “crossing”, “cross pollination”or “cross-breeding” refer to the process by which the pollen of oneflower on one plant is applied (artificially or naturally) to the ovule(stigma) of a flower on another plant. Backcrossing is a process inwhich a breeder repeatedly crosses hybrid progeny, for example a firstgeneration hybrid (F1), back to one of the parents of the hybridprogeny. Backcrossing can be used to introduce one or more single locusconversions from one genetic background into another.

In some embodiments, the present disclosure provides methods forobtaining plant genotypes. As used herein, the term “genotype” refers tothe genetic makeup of an individual cell, cell culture, tissue, organism(e.g., a plant), or group of organisms.

In some embodiments, the present disclosure provides homozygotes. Asused herein, the term “homozygote” refers to an individual cell or planthaving the same alleles at one or more loci.

In some embodiments, the present disclosure provides homozygous plants.As used herein, the term “homozygous” refers to the presence ofidentical alleles at one or more loci in homologous chromosomalsegments.

In some embodiments, the present disclosure provides hemizygotes. Asused herein, the term “hemizygotes” or “hemizygous” refers to a cell,tissue, organism or plant in which a gene is present only once in agenotype, as a gene in a haploid cell or organism, a sex-linked gene inthe heterogametic sex, or a gene in a segment of chromosome in a diploidcell or organism where its partner segment has been deleted.

In some embodiments, the present disclosure provides heterozygotes. Asused herein, the terms “heterozygote” and “heterozygous” refer to adiploid or polyploid individual cell or plant having different alleles(forms of a given gene) present at least at one locus. In someembodiments, the cell or organism is heterozygous for the gene ofinterest that is under control of the synthetic regulatory element.

The disclosure provides self-pollination populations. As used herein,the term “self-crossing”, “self-pollinated” or “self-pollination”,“self-fertilized” or “self-fertilization” means the pollen of one floweron one plant is applied (artificially or naturally) to the ovule(stigma) of the same or a different flower on the same plant. In someembodiments, plants of the present disclosure are genetically stable,such that pollination between plants of the same cultivar producesoffspring are still considered part of the same cultivar.

In some embodiments, the present disclosure teaches cannabis plants,which are an annual, dioecious, flowering herb. Its leaves are typicallypalmately compound or digitate, with serrated leaflets. Cannabisnormally has imperfect flowers, with staminate “male” and pistillate“female” flowers occurring on separate plants. It is not unusual,however, for individual plants of some cannabis varieties to separatelybear both male and female flowers (i.e., have monoecious plants).Although monoecious plants are often referred to as “hermaphrodites,”true hermaphrodites (which are less common in cannabis) bear staminateand pistillate structures on individual flowers, whereas monoeciousplants bear male and female flowers at different locations on the sameplant. In some embodiments, plants of the ‘05.09.24.S1’ variety havebeen feminized, and only produce female inflorescences. In someembodiments, seeds of the ‘05.09.24.S1’ variety produce plants that aregreater than 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% female.

Persons having skill in the art will be familiar with ways of inducingmale flowers in otherwise female plants, including rodelization orcolloidal silver treatments. Briefly, rodelization is the process ofstressing female plants to induce pollen sac formations. This can bedone by allowing unfertilized female flowers to go beyond harvestmaturity in flowering conditions, which will trigger the formation ofpollen sacs in the plant's last effort to self-fertilize before the endof the life cycle. Another way of triggering the formation pollen inotherwise feminized plants is to spray the feminized plants at theflowering stage with colloidal silver solutions (e.g. >30 ppm). Afterseveral sprays, the plants will start forming pollen sacks. Other formsof silver, such as silver nitrate and silver thiosulfate are alsoeffective. Also, hormones such as gibberellins can be used to inducemale flowers on female cannabis plants. Additional methods of inducingmale flowers have been known to one of ordinary skill in the art, e.g.,methods discussed in Ram and Sett (Theoretical and Applied Genetics,1982, 62(4):369-375) and methods discussed in Ram and Jaiswal (Plant,1972, 105(3):263-266), each of which is incorporated by reference in itsentirety for all purposes.

As used herein, a “dioecious” plant refers to a plant having either onlymale flowers (androecious) or female flowers (gynoecious).

As used herein, a “monoecious” plant is a plant having both male andfemale or bisexual flowers, or both female and male or bisexual flowers.Plants bearing separate flowers of both sexes at the same time arecalled simultaneously or synchronously monoecious. Plants bearingflowers of one sex at one time are called consecutively monoecious.

The disclosure provides ovules and pollens of plants. As used hereinwhen discussing plants, the term “ovule” refers to the femalegametophyte, whereas the term “pollen” means the male gametophyte.

The disclosure provides methods for obtaining plants comprisingrecombinant genes through transformation. As used herein, the term“transformation” refers to the transfer of nucleic acid (i.e., anucleotide polymer) into a cell. As used herein, the term “genetictransformation” refers to the transfer and incorporation of DNA,especially recombinant DNA, into a cell.

The disclosure provides transformants comprising recombinant genes. Asused herein, the term “transformant” refers to a cell, tissue ororganism that has undergone transformation. The original transformant isdesignated as “T0.” Selfing the T0 produces a first transformedgeneration designated as “F1” or “T1.”

In some embodiments, the present disclosure refers to inflorescencesfrom a cannabis plant comprising particular cannabinoid and terpenecontents (i.e., inflorescences comprising no more than 0.3% THC). Insome embodiments, inflorescences, such as dried inflorescences,described as having cannabinoid content are female inflorescences. Insome embodiments, the inflorescences are grown “sinsemilla,” in theabsence of male plants to avoid pollination. Thus in some embodiments,the female inflorescences of the present disclosure are seedless, and inmany cases, un-pollinated. The term “inflorescence” and “flower” areused interchangeably throughout this document.

Unless otherwise noted, references to cannabinoids in a plant, plantpart, extract, or composition of the present disclosure should beunderstood as references to both the acidic and decarboxylated versionsof the compound (e.g., potential THC as determined by the conversionguidelines described in this document, and understood by those skilledin the art). For example, unless otherwise stated or clear from thecontext, references to high CBD contents of a cannabis plant in thisdisclosure should be understood as references to the combined CBD andCBDA content (accounting for weight loss during decarboxylation).

Detailed Botanical Description

The present disclosure relates to a new and distinct hemp (Cannabissativa L.) cultivar designated as ‘05.09.24.S1.’ Whole-plant hempextracts from ‘05.09.24.S1’ contain an assortment of phytocannabinoids(e.g., CBD), terpenes, flavonoids and other minor but valuable hempcompounds that work synergistically to heighten effects of productsproduced from ‘05.09.24.S1.’ This synergistic effect is sometimesreferred to as the “entourage effect.” ‘05.09.24.S1’ extracts can beused to produce a variety of products, including liquid and capsuleforms for oral administration, topical products, cosmetic products,infused beverages, sport products and hemp-infused pet treats.

Despite cannabis being consumed since at least the third millennium BC,complete scientific corroboration for uses of CBD are still in theirinfancy. Industry reports suggest CBD is used for a variety of healthand wellness purposes, including as a sleep aid, coping with dailystress, fighting anxiety, relieving pain, assisting with cognitivefunction and boosting immune health. Significant research is currentlybeing conducted at a variety of laboratories on the use of CBD as itrelates to epilepsy, Post-Traumatic Stress Disorder (PTSD), cancer,autism, neuroprotection, anti-inflammatory effects, anti-tumor effectsand anti-psychotic effects.

‘05.09.24.S1’ is a selection resulting from a series ofcontrolled-crosses using proprietary cannabis plant(s). Specifically,‘05.09.24.S1’ was propagated by seed from the self-fertilization of aparent cannabis plant named ‘BIHEMP 050924’ (USPP32,473) at applicant'sfacilities. The primary goal of the breeding program was to develop anew hemp variety with improved disease resistance, as well as highcannabidiolic acid (CBDA) concentrations and low tetrahydrocannabinolicacid (THCA) concentrations in its mature female flowers.

‘05.09.24.S1’ has not been observed under all possible environmentalconditions, and the phenotype may vary significantly with variations inenvironment. The following observations, measurements, and comparisonsdescribe this plant as grown in a non-heated greenhouse structure inSalinas, Calif.

Plants for the botanical measurements in the present application areannual plants. In the following description, the color determination isin accordance with The Royal Horticultural Society Colour Chart, fifthEdition (2007), except where general color terms of ordinary dictionarysignificance are used.

Breeding History of the Female Parent.

A pedigree of the hemp plant named ‘05.09.24.S1’ is presented in FIG. 1.

The lineage of proprietary female parent ‘BIHEMP 050924’ comprises fourgenerational crossings. The first crossing was made between aproprietary parental female plant with unknown genealogy (THV01) with aparental pollen donor plant with unknown genealogy (CDB05.S1-P24). Aresulting F1 progeny line (V24) was produced.

For the second crossing, the germinated plants from the V24 line wasself-fertilized to produce an F2 generation (V24.S1). Of the resultingoffspring, one progeny plant (V24.S1.N5) was selected as a pollenacceptor and another progeny plant (V24.S1.O3) was chosen as a pollendonor.

For the third crossing, a single F₂ Female plant (V24.S1.N5) was chosenand crossed with its sibling F₂ pollen donor plant (V24.S1.O3) togenerate an F3 generation (O3.N5).

The fourth generation (05.09.24) was produced by crossing two siblings,O3.N5.05 (pollen acceptor) and O3.N5.09 (pollen donor). Out of the F4offspring, all F4 plants exhibited pistillate flowers with resistance topests and diseases, and showed high vigor throughout the entirelifecycle. Of these F4 females, the healthiest and most vigorousindividual was chosen to be ‘BIHEMP 050924’.

Self-Crossing of Induced Female Parental Line. The F4 female parent‘BIHEMP 050924’ was treated to trigger the formation of intersexual andfully altered male flowers on the newly formed primary lateral branches,thus making self-fertilization possible within the ‘BIHEMP 050924’plants. The induced ‘BIHEMP 050924’ female plants formed the maleflowers, and were consequently self-pollinated to develop seeds. Thisprocess was repeated more than three times. The resulting seeds aftermultiple rounds of selfing produce a novel, uniform, genetically stable,and feminized plant designated as ‘05.09.24.S1’. For the germinationtest, 400 seeds of ‘05.09.24.S1’ cultivar were prechilled for 3 days at10° C. and placed for 10 days at 20° C.-30° C. with a moisturizedsetting. 87% of the seeds were germinated and viable. All the observed‘05.09.24.S1’ plants grown from the seeds displayed a very upright plantform from main axis dominance. Also, all 93 plants tested for sexingwere female plants.

The primary selection criteria for the new and distinct hemp cultivardisclosed herein is as follows: (i) Phenotype-Structure, (ii)/Phenolic(s) Scoring, (iii) Resistance/Susceptibility to Pest andDisease; (iv) Chemotypic Analysis for Cannabinoids, Terpenes and/orother secondary metabolites.

‘05.09.24.S1’ was initially propagated and is maintained by seed.

Tables 1-5, below, provide the morphological and physiologicalcharacteristics of the ‘05.09.24.S1’ variety. A minimum of 25 plantswere measured when the plants were 100 days old (i.e. 15 days inpropagation stage, 25 days in vegetative stage, and 60 days in floweringstage). That is, the ‘05.09.24.S1’ plants were observed at the peak ofthe floral maturity right before harvest. Morphological andphysiological characteristics of ‘BIHEMP 050924’ gathered in the samelocation when the plants were 100 days old for comparative purposes.

TABLE 1 General Characteristics New Variety Check Variety Characteristic(05.09.24.S1) (BIHEMP 050924) Plant life forms An herbaceous plant(herb) An herbaceous plant (herb) Plant growth habit An upright,tap-rooted annual An upright, tap-rooted annual plant; forming fibrousroots plant; forming fibrous roots when asexually propagated whenasexually propagated Plant origin A controlled self-cross of Acontrolled-cross between BIHEMP 050924 (O3.N5.05) pollen acceptor(O3.N5.05) and pollen donor (O3.N5.09) Plant propagation Propagated byseed Asexually propagated by stem cuttings and cloning Propagation easeEasy Easy Plant Height 1.5 m-2.5 m 0.3 m-1.2 m (Unit: m) Plant Width 50cm-128 cm 56 cm-90 cm (Unit: cm) Plant vigor High High Resistance topests Resistant to pests as follows: Resistant to pests as follows: ordiseases (1) Two-spotted spider (1) Two-spotted spider mite Tetranychusmite Tetranychus urticae (Koch) urticae (Koch) (2) Aphid species suchas: (2) Aphids species such as: Cannabis Aphid (Phorodon Cannabis Aphid(Phorodon cannabis); cannabis); Green Peach Aphid (Myzus Green PeachAphid (Myzus persicae (Sulzer)); persicae (Sulzer)); Foxglove Aphid(Aulacorthum Foxglove Aphid (Aulacorthum solani); solani); Peach Aphid(Macrosiphum Peach Aphid (Macrosiphum euphorbiae); euphorbiae); BlackBean Aphid (Aphis Black Bean Aphid (Aphis fabae) fabae) (3) Whitefly(Trialeurodes (3) Whitefly (Trialeurodes vaporariorum) vaporariorum) (4)Lepidoptera species such (4) Lepidoptera species such as: as: Armyworm(Spodoptera Armyworm (Spodoptera frugiperda); Cabbage White frugiperda);Cabbage White (Pieris rapae); Painted (Pieris rapae); Painted Lady(Vanessa Lady (Vanessa cardui); Lepidoptera sp. cardui); Lepidoptera sp.Resistant to diseases as follows; Resistant to diseases as PowderyMildew (Podosphaera follows; Powdery xanthii) Mildew (Podosphaeraxanthii) Time to Harvest 9-11 weeks 9-10 weeks Genetically- No Nomodified organism

TABLE 2 Leaf/Foliage New Variety Check Variety Characteristic(05.09.24.S1) (BIHEMP 050924) Leaf structure Lanceolate leaflet bladeswith Lanceolate leaflet glandular hairs blades with glandular hairs Leafshape Palmately Compound Palmately Compound Leaf arrangement Opposite atseedling/immature Opposite at seedling/ (Phyllotaxy) stage; immaturestage; Alternate at mature/flowering Alternate at mature/ stageflowering stage Leaf margin Dentate, coarsely serrated, and Dentate,coarsely the teeth point towards the tip serrated, and the teeth pointtowards the tip Leaf hair Leaf hairs occur on both the Leaf hairs occurupper and lower surface(s) of on both the upper the leaves and lowersurface(s) of the leaves Leaf length with 25.60 cm-26.10 cm 19.50cm-21.30 cm petiole (Unit: cm) (average 25.90 cm) (average 20.40 cm) No.of leaflets 5-7 5-7 Middle largest 18.10 cm-18.30 cm 10.20 cm-13.50 cm(longest) leaflet (average 18.20 cm) (average 12.10 cm) length (Unit:cm) Middle largest 3.80 cm-4.50 cm 1.70 cm-2.50 cm (longest) leaflet(average 4.10 cm) (average 2.20 cm) width* (Unit: cm) Middle largestAbout 18.2:4.1 About 12.1:2.2 (longest) leaflet length/width ratio No.teeth of middle 32-39 19-25 leaflet (secondary teeth form on primaryteeth in reference to leaf margin) Leaf (upper side - 135A 139B adaxial)color (RHS No.) Leaf (lower side - 130D 139C abaxial) color (RHS No.)Leaf glossiness Medium Strong Vein/midrib shape Obliquely continuousObliquely continuous throughout leaflet throughout leaflet Vein/midribcolor 154C 151D (RHS No.) Petiole length* 7.50 cm-7.70 cm 6.50 cm-8.40cm (Unit: cm) (average 7.60 cm) (average 7.70 cm) Petiole color 142B153A (RHS No.) Petiole Present Present anthocyanin coloration Intensityof Petiole Very Weak Moderate anthocyanin Stipule shape EllipticalElliptical Stipule length 0.90 cm-1.50 cm 0.49 cm-0.66 cm (Unit: cm)(average 1.27 cm) (average 0.60 cm) Stipule color 134A 134A (RHS No.)

TABLE 3 Stem New Variety Check Variety Characteristic (05.09.24.S1)(BIHEMP 050924) Stem shape Hollow, ribbed, textured Hollow, ribbed,textured Stem diameter at 1.80 cm-3.40 cm 2.60 cm-3.10 cm base (Unit:cm) (average 2.60 cm) (average 2.80 cm) Stem color N144C 149D (RHS No.)Stem pith type Thick Thick Depth of main stem Deep Medium ribs/groovesInternode length 5.20 cm-5.80 cm 4.20 cm-4.85 cm (Average 5.43 cm)(average 4.53 cm)

TABLE 4 Inflorescence (Female/Pistillate Flowers) New Variety CheckVariety Characteristic (05.09.24.S1) (BIHEMP 050924) FloweringPistillate flowers emerge in an Pistillate flowers emerge in an(blooming) habit upward fashion from apical upward fashion from apicalmeristems meristems Inflorescence Flowers develop above the Flowersdevelop above the position relative to apical portions of main andapical portions of main and foliage lateral axes lateral axes FlowerRaceme-like cyme; Raceme-like cyme; arrangement Compound in natureCompound in nature Number of flowers 40-95 per cyme; 45-90 per cyme; perspike, panicle 14-21 panicles per plant 8-12 panicles per plant orraceme Flower shape Urceolate in shape; Urceolate in shape; Compoundclusters borne in Compound clusters borne in racemes; racemes; Eachindividual flower has a Each individual flower has a small green bractenclosing an small green bract enclosing an ovary with two-long, slenderovary with two-long, slender stigmas projecting well above stigmasprojecting well above the bract the bract Flower (individual 10.15mm-11.55 mm 9.22 mm-10.10 mm pistillate) length (average 10.85 mm)(average 9.74 mm) (Unit: mm) Flower (compound 8.40-9.10 cm 6.0 cm-9.1 cmcyme) diameter (average 8.80 cm) (average 7.77 cm) (Unit: cm) Corollashape N/A N/A Corolla color N/A N/A (RHS No.) Bract shape UrceolateUrceolate Bract size/length 3.50 mm-4.40 mm 2.90 mm-3.40 mm (Unit: cm ormm) (average 4.00 mm) (average 3.17 mm) Bract color 134A 134A (RHS No.)Stigma shape Pointed, linear Pointed, linear Stigma length 1.9 mm-2.3 mm1.3 mm-1.7 mm (Unit: mm) (average 2.13 mm) (average 1.50 mm) Stigmacolor 44A 44A (RHS No.) Calyx shape No defined calyx No defined calyx(general description) Calyx color N/A N/A (RHS No.) Trichome shapeCapitate sessile trichomes, Capitate sessile trichomes, which arepresent on the which are present on the leaves leaves of plants as wellas of plants as well as being being noticed in the flowers; noticed inthe flowers; Capitate stalked trichomes are Capitate stalked trichomesare present in the flowers; present in the flowers; Bulbous andnon-glandular Bulbous and non-glandular trichomes are also present,trichomes are also present, most most noticeable on the noticeable onthe petioles, petioles, stems, and leaves. stems, and leaves. Trichomecolor Capitate sessile trichomes Capitate sessile trichomes 157A (RHSNo.) 157A (early flower - at day 40 (early flower - at day 40 of offlowering) flowering) Capitate stalked trichomes Capitate stalkedtrichomes N30B (late flowering - at day N30B (late flowering - at day 55to day 70 of flowering) 55 to day 70 of flowering) Bulbous andnon-glandular Bulbous and non-glandular trichomes trichomes 157A 157ATerminal bud shape Oblong Oblong Terminal bud 134B 134B color (RHS No.)Pedicel (Presence Present Present or absence) Pedicel color 147D 150D(RHS No.) Sepal Absent Absent Sepal color N/A N/A (RHS No.) PetalAbsent; Apetulous Absent; Apetulous Petal color N/A N/A (RHS No.)Staminate flower N/A N/A Pollen N/A N/A Seed shape Globular and texturedGlobular and textured Seed size/length 3.10 mm-3.90 mm 2.00 mm-2.30 mm(average 3.43 mm) (average 2.13 mm) Seed color 199A 199A (RHS No.)Marbling of seed Medium Weak N/A: Not Available

TABLE 5 Other Characteristics New Variety Check Variety Characteristic(05.09.24.S1) (BIHEMP 050924) Aroma Sweet yet savory; mouth Pungent, yetsweet watering Proportion of Null to low % when grown Null to low % whengrown Hermaphrodite under ideal environmental under ideal environmentalconditions conditions Time period and 50-70 days 60-70 days condition offlowering/blooming (Unit: days or weeks) Plant Hardiness Hardy (20° F.to 120° F.) Hardy (20° F. to 120° F.) Breaking action Flexible, highlyresistant to Flexible, highly resistant to breakage breakage Rootingrate after 99% under ideal temperature 99% under ideal temperaturecutting/cloning and relative humidity and relative humidity (undercertain or specific condition) Types of Cutting Meristem Meristem forCloning (stem, leaf, root etc.) Shipping quality High/great High/greatStorage life 3-8 months with minor 3-8 months with minor change changein metabolites, in metabolites, smell/taste, smell/taste, and/orphysical and/or physical appearance appearance Minor decrease in greenMinor decrease in green coloration coloration Market use of Dryinhalable extract(s) Dry inhalable extract(s) flower Productivity of0.5-1.5 pounds per plant 0.5-1.0 pounds per plant flower (weight perplant) Terpenes See Table 7. See Table 7. Cannabinoids About 0.00% toabout 0.26% About 0.21% to about 0.43% THC potential by dry weight THCpotential by dry weight of of the inflorescence. the inflorescence.Median of about 4.39% About 5.02% to about 10.86% potential by dryweight of the CBD potential by dry weight of inflorescence. theinflorescence.

Table 6 includes detailed information of the hemp plant named‘05.09.24.S1’ for profiles of total CBD and THC concentrations as testedon one hundred seventy eight flower samples. The hemp plant has beentested in a laboratory setting and/or facility to determine cannabinoidsand terpenes concentrations in the hemp plant named ‘05.09.24.S1’according to the procedures provided in Giese et al. (Journal of AOACInternational (2015) 98(6):1503-1522). Table 7 includes detailedinformation about the terpene profile of the hemp plant named‘05.09.24.S1’.

As used herein, the term “maturity,” “harvest maturity,” or “floralmaturity” refers to the developmental stage at which the ‘05.09.24.S1’plant is harvested. Persons having skill in the art will recognizematurity based on the plant's morphologies. It is also good practice toconduct periodic cannabinoid content (i.e., potency) tests throughoutthe development of the plant to ensure that harvest occurs at maturity.Since ‘05.09.24.S1’ is a photoperiod sensitive plant, time to maturitydepends largely on the day/night cycles where grown.

In some embodiments, stigma color can also be an indication of maturity.For example, in some embodiments, if all stigmas are red or browning itcould indicate ‘05.09.24.S1’ is past maturity. In some embodiments, itis preferable to harvest when some stigmas are still white.

Growing conditions throughout the plant's life cycle, nutrientvariations, and environmental factors can all influence the amount oftime for ‘05.09.24.S1’ plants to reach harvest maturity. The presentdisclosure uses the terms “maturity,” “harvest maturity,” and “floralmaturity” interchangeably. In some embodiments, harvest maturity canencompass any period after the emergence of inflorescences, but beforethe THC content of any inflorescence surpasses 0.3%.

In some instances, the botanical descriptions disclosed herein reflectthe range of phenotypical variation observed under indoor and outdoorgrowth conditions. Total Potential THC/CBD contents presented in thisdocument reflect the total potential (i.e., decarboxylated) THC and CBDcontent after decarboxylation of the THCA and CBDA contents of thesample. The formula used for this calculation is reproduced below forthe Office's convenience. Total THC=THC+(THCA*(0.877)). TotalCBD=CBD+(CBDA*(0.877)). Additionally, CBGA can be converted to activeCBG by multiplying 87.8% to CBGA. Thus, the maximum amount of CBG is:CBG_(max)=(CBGA×0.878)+CBG.

When ‘05.09.24.S1’ is compared to the check variety ‘BIHEMP 050924’,‘05.09.24.S1’ and ‘BIHEMP 050924’ were both bred primarily for pestand/or disease resistance. Common pests that ‘05.09.24. S1’ and ‘BIHEMP050924’ are resistant/tolerant to include: Two-spotted spider mite,Whitefly, Aphid species (such as Cannabis Aphid; Green Peach Aphid;Foxglove Aphid; Peach Aphid; Black Bean Aphid), and Lepidoptera species(such as Armyworm; Cabbage White; Painted Lady; Lepidoptera sp.)

‘05.09.24.S1’ and ‘BIHEMP 050924’ show high vigor and health.‘05.09.24.S1’ is taller and wider in plant height and width than ‘BIHEMP050924’ in maturity. Also, ‘05.09.24.S1’ is a week or two shorter than‘BIHEMP 050924’ in flowering time period and has more panicles withlarger inflorescences. The aroma of ‘05.09.24.S1’ is sweet, yet savorywith mouth watering, while that of ‘BIHEMP 050924’ is pungent, yetsweet.

Cannabis Hemp Breeding Methods

In some embodiments, the plants of the present disclosure can be used toproduce new plant varieties. In some embodiments, the plants are used todevelop new, unique and superior varieties or hybrids with desiredphenotypes. As used herein, the term “plant breeding techniques”comprises all of the plant breeding techniques disclosed in this sectionof the application, and well known to persons having skill in the art.Thus, in some embodiments, plant breeding methods encompass theapplication of recurrent selection, mass selection, hybridization,open-pollination, backcrossing, pedigree breeding, marker assistedselection breeding, mutation breeding, gene editing, and combinationsthereof.

In some embodiments, selection methods, e.g., molecular marker assistedselection, can be combined with breeding methods to accelerate theprocess. Additional breeding methods have been known to one of ordinaryskill in the art, e.g., methods discussed in Chahal and Gosal(Principles and procedures of plant breeding: biotechnological andconventional approaches, CRC Press, 2002, ISBN 084931321X,9780849313219), Taji et al. (In vitro plant breeding, Routledge, 2002,ISBN 156022908X, 9781560229087), Richards (Plant breeding systems,Taylor & Francis US, 1997, ISBN 0412574500, 9780412574504), Hayes(Methods of Plant Breeding, Publisher: READ BOOKS, 2007, ISBN1406737062,9781406737066), each of which is incorporated by reference in itsentirety for all purposes. The Cannabis genome has been sequencedrecently (van Bakel et al., The draft genome and transcriptome ofCannabis sativa, Genome Biology, 12(10):R102, 2011). Molecular markersfor cannabis plants are described in Datwyler et al. (Genetic variationin hemp and marijuana (Cannabis sativa L.) according to amplifiedfragment length polymorphisms, J Forensic Sci. 2006 March; 51(2):371-5),Pinarkara et al., (RAPD analysis of seized marijuana (Cannabis sativaL.) in Turkey, Electronic Journal of Biotechnology, 12(1), 2009), Hakkiet al., (Inter simple sequence repeats separate efficiently hemp frommarijuana (Cannabis sativa L.), Electronic Journal of Biotechnology,10(4), 2007), Datwyler et al., (Genetic Variation in Hemp and Marijuana(Cannabis sativa L.) According to Amplified Fragment LengthPolymorphisms, J Forensic Sci, March 2006, 51(2):371-375), Gilmore etal. (Isolation of microsatellite markers in Cannabis sativa L.(marijuana), Molecular Ecology Notes, 3(1):105-107, March 2003),Pacifico et al., (Genetics and marker-assisted selection of chemotype inCannabis sativa L.), Molecular Breeding (2006) 17:257-268), and Mendozaet al., (Genetic individualization of Cannabis sativa by a short tandemrepeat multiplex system, Anal Bioanal Chem (2009) 393:719-726), each ofwhich is herein incorporated by reference in its entirety for allpurposes.

In some embodiments, molecular markers are designed and made, based onthe genome of the plants of the present application. In someembodiments, the molecular markers are selected from IsozymeElectrophoresis, Restriction Fragment Length Polymorphisms (RFLPs),Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily PrimedPolymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting(DAF), Sequence Characterized Amplified Regions (SCARs). AmplifiedFragment Length Polymorphisms (AFLPs), and Simple Sequence Repeats(SSRs) which are also referred to as Microsatellites, etc. Methods ofdeveloping molecular markers and their applications are described byAvise (Molecular markers, natural history, and evolution, Publisher:Sinauer Associates, 2004, ISBN 0878930418, 9780878930418), Srivastava etal. (Plant biotechnology and molecular markers, Publisher: Springer,2004, ISBN1402019114, 9781402019111), and Vienne (Molecular markers inplant genetics and biotechnology, Publisher: Science Publishers, 2003),each of which is incorporated by reference in its entirety for allpurposes.

The molecular markers can be used in molecular marker assisted breeding.For example, the molecular markers can be utilized to monitor thetransfer of the genetic material. In some embodiments, the transferredgenetic material is a gene of interest, such as genes that contribute toone or more favorable agronomic phenotypes when expressed in a plantcell, a plant part, or a plant.

Details of existing cannabis plants varieties and breeding methods aredescribed in Potter et al. (2011, World Wide Weed: Global Trends inCannabis Cultivation and Its Control), Holland (2010, The Pot Book: AComplete Guide to Cannabis, Inner Traditions/Bear & Co, ISBN1594778981,9781594778988), Green I (2009, The Cannabis Grow Bible: The DefinitiveGuide to Growing Marijuana for Recreational and Medical Use, Green CandyPress, 2009, ISBN 1931160589, 9781931160582), Green II (2005, TheCannabis Breeder's Bible: The Definitive Guide to Marijuana Genetics,Cannabis Botany and Creating Strains for the Seed Market, Green CandyPress, 1931160279, 9781931160278), Starks (1990, Marijuana Chemistry:Genetics, Processing & Potency, ISBN 0914171399, 9780914171393), Clarke(1981, Marijuana Botany, an Advanced Study: The Propagation and Breedingof Distinctive Cannabis, Ronin Publishing, ISBN 091417178X,9780914171782), Short (2004, Cultivating Exceptional Cannabis: An ExpertBreeder Shares His Secrets, ISBN 1936807122, 9781936807123), Cervantes(2004, Marijuana Horticulture: The Indoor/Outdoor Medical Grower'sBible, Van Patten Publishing, ISBN 187882323X, 9781878823236), Franck etal. (1990, Marijuana Grower's Guide, Red Eye Press, ISBN 0929349016,9780929349015), Grotenhermen and Russo (2002, Cannabis and Cannabinoids:Pharmacology, Toxicology, and Therapeutic Potential, Psychology Press,ISBN 0789015080, 9780789015082), Rosenthal (2007, The Big Book of Buds:More Marijuana Varieties from the World's Great Seed Breeders, ISBN1936807068, 9781936807062), Clarke, R C (Cannabis: Evolution andEthnobotany 2013 (In press)), King, J (Cannabible Vols 1-3, 2001-2006),and four volumes of Rosenthal's Big Book of Buds series (2001, 2004,2007, and 2011), each of which is herein incorporated by reference inits entirety for all purposes.

Classical breeding methods can be included in the present disclosure tointroduce one or more recombinant expression cassettes of the presentdisclosure into other plant varieties, or other close-related speciesthat are compatible to be crossed with the transgenic plant. In someembodiments, the recombinant expression cassette can encode for adesirable phenotype, including herbicide resistance, disease or pestresistance, insect resistance, resistance to antibiotics, or additionaltraits, as disclosed in this application.

In some embodiments, said method comprises (i) crossing any one of theplants of the present disclosure comprising the expression cassette as adonor to a recipient plant line to create a F1 population; (ii)selecting offspring that have expression cassette. Optionally, theoffspring can be further selected by testing the expression of the geneof interest. Thus in some embodiments, the present disclosure teachescrossing a transgenic plant with the presently disclosed ‘05.09.24.S1’plant.

In some embodiments, complete chromosomes of the donor plant aretransferred. For example, the transgenic plant with the expressioncassette can serve as a male or female parent in a cross pollination toproduce offspring plants, wherein by receiving the transgene from thedonor plant, the offspring plants have the expression cassette.

In a method for producing plants having the expression cassette,protoplast fusion can also be used for the transfer of the transgenefrom a donor plant to a recipient plant. Protoplast fusion is an inducedor spontaneous union, such as a somatic hybridization, between two ormore protoplasts (cells in which the cell walls are removed by enzymatictreatment) to produce a single bi- or multi-nucleate cell. The fusedcell that may even be obtained with plant species that cannot beinterbred in nature is tissue cultured into a hybrid plant exhibitingthe desirable combination of traits. More specifically, a firstprotoplast can be obtained from a plant having the expression cassette.A second protoplast can be obtained from a second plant line, optionallyfrom another plant species or variety, preferably from the same plantspecies or variety, that comprises commercially desirablecharacteristics, such as, but not limited to disease resistance, insectresistance, valuable grain characteristics (e.g., increased seed weightand/or seed size) etc. The protoplasts are then fused using traditionalprotoplast fusion procedures, which are known in the art to produce thecross.

Alternatively, embryo rescue may be employed in the transfer of theexpression cassette from a donor plant to a recipient plant. Embryorescue can be used as a procedure to isolate embryos from crosseswherein plants fail to produce viable seed. In this process, thefertilized ovary or immature seed of a plant is tissue cultured tocreate new plants (see Pierik, 1999, In vitro culture of higher plants,Springer, ISBN 079235267x, 9780792352679, which is incorporated hereinby reference in its entirety).

In some embodiments, the recipient plant is an elite line having one ormore certain desired traits. Examples of desired traits include but arenot limited to those that result in increased biomass production,production of specific chemicals, increased seed production, improvedplant material quality, increased seed oil content, etc. Additionalexamples of desired traits include pest resistance, vigor, developmenttime (time to harvest), enhanced nutrient content, novel growthpatterns, aromas or colors, salt, heat, drought and cold tolerance, andthe like. Desired traits also include selectable marker genes (e.g.,genes encoding herbicide or antibiotic resistance used only tofacilitate detection or selection of transformed cells), hormonebiosynthesis genes leading to the production of a plant hormone (e.g.,auxins, gibberellins, cytokinins, abscisic acid and ethylene that areused only for selection), or reporter genes (e.g. luciferase,β-glucuronidase, chloramphenicol acetyl transferase (CAT, etc.). Therecipient plant can also be a plant with preferred chemicalcompositions, e.g., compositions preferred for medical use or industrialapplications.

Classical breeding methods can be used to produce new varieties ofcannabis according to the present disclosure. Newly developed F1 hybridscan be reproduced via asexual reproduction.

Open-Pollinated Populations. The improvement of open-pollinatedpopulations of such crops as cannabis, rye, many maizes and sugar beets,herbage grasses, legumes such as alfalfa and clover, and tropical treecrops such as cacao, coconuts, oil palm and some rubber, dependsessentially upon changing gene-frequencies towards fixation of favorablealleles while maintaining a high (but far from maximal) degree ofheterozygosity. Uniformity in such populations is impossible andtrueness-to-type in an open-pollinated variety is a statistical featureof the population as a whole, not a characteristic of individual plants.Thus, the heterogeneity of open-pollinated populations contrasts withthe homogeneity (or virtually so) of inbred lines, clones and hybrids.

Population improvement methods fall naturally into two groups, thosebased on purely phenotypic selection, normally called mass selection,and those based on selection with progeny testing. Interpopulationimprovement utilizes the concept of open breeding populations; allowinggenes to flow from one population to another. Plants in one population(cultivar, strain, ecotype, or any germplasm source) are crossed eithernaturally (e.g., by wind) or by hand or by bees (commonly Apis melliferaL. or Megachile rotundata F.) with plants from other populations.Selection is applied to improve one (or sometimes both) population(s) byisolating plants with desirable traits from both sources.

There are basically two primary methods of open-pollinated populationimprovement. First, there is the situation in which a population ischanged en masse by a chosen selection procedure. The outcome is animproved population that is indefinitely propagatable by random-matingwithin itself in isolation. Second, the synthetic variety attains thesame end result as population improvement but is not itself propagatableas such; it has to be reconstructed from parental lines or clones. Theseplant breeding procedures for improving open-pollinated populations arewell known to those skilled in the art and comprehensive reviews ofbreeding procedures routinely used for improving cross-pollinated plantsare provided in numerous texts and articles, including: Allard,Principles of Plant Breeding, John Wiley & Sons, Inc. (1960); Simmonds,Principles of Crop Improvement, Longman Group Limited (1979); Hallauerand Miranda, Quantitative Genetics in Maize Breeding, Iowa StateUniversity Press (1981); and, Jensen, Plant Breeding Methodology, JohnWiley & Sons, Inc. (1988).

Mass Selection. In mass selection, desirable individual plants arechosen, harvested, and the seed composited without progeny testing toproduce the following generation. Since selection is based on thematernal parent only, and there is no control over pollination, massselection amounts to a form of random mating with selection. As statedherein, the purpose of mass selection is to increase the proportion ofsuperior genotypes in the population.

Mutation breeding is another method of introducing new traits into thehemp plants of the present disclosure. Mutations that occurspontaneously or are artificially induced can be useful sources ofvariability for a plant breeder. The goal of artificial mutagenesis isto increase the rate of mutation for a desired characteristic. Mutationrates can be increased by many different means including temperature,long-term seed storage, tissue culture conditions, radiation; such asX-rays, Gamma rays (e.g., cobalt 60 or cesium 137), neutrons, (productof nuclear fission by uranium 235 in an atomic reactor), Beta radiation(emitted from radioisotopes such as phosphorus 32 or carbon 14), orultraviolet radiation (preferably from 2500 to 2900 nm), or chemicalmutagens (such as base analogues (5-bromo-uracil)), related compounds(8-ethoxy caffeine), antibiotics (streptonigrin), alkylating agents(sulfur mustards, nitrogen mustards, epoxides, ethyleneamines, sulfates,sulfonates, sulfones, lactones), azide, hydroxylamine, nitrous acid, oracridines. Once a desired trait is observed through mutagenesis thetrait may then be incorporated into existing germplasm by traditionalbreeding techniques. Details of mutation breeding can be found inAllard, Principles of Plant Breeding, John Wiley & Sons, Inc. (1960). Inaddition, mutations created in other hemp plants may be used to producea backcross conversion of hemp plants having all phenotypes of the‘05.09.24.S1’ line while comprising the mutation obtained from the otherhemp plants.

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct gene transfer method, suchas microprojectile-mediated delivery, DNA injection, electroporation,and the like. Additionally, expression vectors are introduced into planttissues by using either microprojectile-mediated delivery with abiolistic device or by using Agrobacterium-mediated transformation.Transformant plants obtained with the protoplasm of the subject hempplants are intended to be within the scope of the embodiments of theapplication.

Synthetics. A synthetic variety is produced by crossing inter se anumber of genotypes selected for good combining ability in all possiblehybrid combinations, with subsequent maintenance of the variety by openpollination. Whether parents are (more or less inbred) seed-propagatedlines, makes no difference in principle. Parents are selected on generalcombining ability, sometimes by test crosses or topcrosses, moregenerally by polycrosses. Parental seed lines may be deliberately inbred(e.g. by selfing or sib crossing). However, even if the parents are notdeliberately inbred, selection within lines during line maintenance willensure that some inbreeding occurs. Clonal parents will, of course,remain unchanged and highly heterozygous.

Whether a synthetic can go straight from the parental seed productionplot to the farmer or must first undergo one or two cycles ofmultiplication depends on seed production and the scale of demand forseed.

While mass selection is sometimes used, progeny testing is generallypreferred for polycrosses, because of their operational simplicity andobvious relevance to the objective, namely exploitation of generalcombining ability in a synthetic.

The numbers of parental lines or clones that enter a synthetic varywidely. In practice, numbers of parental lines range from 10 to severalhundred, with 100-200 being the average. Broad based synthetics formedfrom 100 or more clones would be expected to be more stable during seedmultiplication than narrow based synthetics.

Pedigreed varieties. A pedigreed variety is a superior genotypedeveloped from selection of individual plants out of a segregatingpopulation followed by propagation and seed increase of self-pollinatedoffspring and careful testing of the genotype over several generations.This is an open pollinated method that works well with naturallyself-pollinating species. This method can be used in combination withmass selection in variety development. Variations in pedigree and massselection in combination are the most common methods for generatingvarieties in self-pollinated crops.

Hybrids. A hybrid is an individual plant resulting from a cross betweenparents of differing genotypes. Commercial hybrids are now usedextensively in many crops, including corn (maize), sorghum, sugar beet,sunflower and broccoli. Hybrids can be formed in a number of differentways, including by crossing two parents directly (single cross hybrids),by crossing a single cross hybrid with another parent (three-way ortriple cross hybrids), or by crossing two different hybrids (four-way ordouble cross hybrids).

Strictly speaking, most individuals in an out breeding (i.e.,open-pollinated) population are hybrids, but the term is usuallyreserved for cases in which the parents are individuals whose genomesare sufficiently distinct for them to be recognized as different speciesor subspecies. Hybrids may be fertile or sterile depending onqualitative and/or quantitative differences in the genomes of the twoparents. Heterosis, or hybrid vigor, is usually associated withincreased heterozygosity that results in increased vigor of growth,survival, and fertility of hybrids as compared with the parental linesthat were used to form the hybrid. Maximum heterosis is usually achievedby crossing two genetically different, highly inbred lines.

Targeting Induced Local Lesions in Genomes (TILLING). Breeding schemesof the present disclosure can include crosses with TILLING® plant lines.TILLING® is a method in molecular biology that allows directedidentification of mutations in a specific gene. TILLING® was introducedin 2000, using the model plant Arabidopsis thaliana. TILLING® has sincebeen used as a reverse genetics method in other organisms such aszebrafish, corn, wheat, rice, soybean, tomato and lettuce. The methodcombines a standard and efficient technique of mutagenesis with achemical mutagen (e.g., Ethyl methanesulfonate (EMS)) with a sensitiveDNA screening-technique that identifies single base mutations (alsocalled point mutations) in a target gene. EcoTILLING is a method thatuses TILLING® techniques to look for natural mutations in individuals,usually for population genetics analysis (see Comai, et al., 2003 ThePlant Journal 37, 778-786; Gilchrist et al. 2006 Mol. Ecol. 15,1367-1378; Mejlhede et al. 2006 Plant Breeding 125, 461-467; Nieto etal. 2007 BMC Plant Biology 7, 34-42, each of which is incorporated byreference hereby for all purposes). DEcoTILLING is a modification ofTILLING® and EcoTILLING which uses an inexpensive method to identifyfragments (Garvin et al., 2007, DEco-TILLING: An inexpensive method forSNP discovery that reduces ascertainment bias. Molecular Ecology Notes7, 735-746). More detailed description on methods and compositions onTILLING® can be found in U.S. Pat. No. 5,994,075, US 2004/0053236 A1, WO2005/055704, and WO 2005/048692, each of which is hereby incorporated byreference for all purposes.

In some embodiments, TILLING® can also be utilized for plants of thecannabis genus including hemp plants. Thus in some embodiments, thebreeding methods of the present disclosure include breeding with one ormore TILLING plant lines with one or more identified mutations.

Gene editing technologies. Breeding and selection schemes of the presentdisclosure can include crosses with plant lines that have undergonegenome editing. In some embodiments, the breeding and selection methodsof the present disclosure are compatible with plants that have beenmodified using any gene and/or genome editing tool, including, but notlimited to: ZFNs, TALENS, CRISPR, and Mega nuclease technologies. Insome embodiments, persons having skill in the art will recognize thatthe breeding methods of the present disclosure are compatible with manyother gene editing technologies. In some embodiments, the presentdisclosure teaches gene-editing technologies can be applied for a singlelocus conversion, for example, conferring hemp plant with herbicideresistance. In some embodiments, the present disclosure teaches that thesingle locus conversion is an artificially mutated gene or nucleotidesequence that has been modified through the use of breeding techniquestaught herein.

In some embodiments, the breeding and selection methods of the presentdisclosure are compatible with plants that have been modified throughZinc Finger Nucleases. Three variants of the ZFN technology arerecognized in plant breeding (with applications ranging from producingsingle mutations or short deletions/insertions in the case of ZFN-1 and-2 techniques up to targeted introduction of new genes in the case ofthe ZFN-3 technique); 1) ZFN-1: Genes encoding ZFNs are delivered toplant cells without a repair template. The ZFNs bind to the plant DNAand generate site specific double-strand breaks (DSBs). The naturalDNA-repair process (which occurs through nonhomologous end-joining,NHEJ) leads to site specific mutations, in one or only a few base pairs,or to short deletions or insertions; 2) ZFN-2: Genes encoding ZFNs aredelivered to plant cells along with a repair template homologous to thetargeted area, spanning a few kilo base pairs. The ZFNs bind to theplant DNA and generate site-specific DSBs. Natural gene repairmechanisms generate site-specific point mutations e.g. changes to one ora few base pairs through homologous recombination and the copying of therepair template; and 3) ZFN-3: Genes encoding ZFNs are delivered toplant cells along with a stretch of DNA which can be several kilo basepairs long and the ends of which are homologous to the DNA sequencesflanking the cleavage site. As a result, the DNA stretch is insertedinto the plant genome in a site-specific manner.

In some embodiments, the breeding and selection methods of the presentdisclosure are compatible with plants that have been modified throughTranscription activator-like (TAL) effector nucleases (TALENs). TALENSare polypeptides with repeat polypeptide arms capable of recognizing andbinding to specific nucleic acid regions. By engineering the polypeptidearms to recognize selected target sequences, the TAL nucleases can beused to direct double stranded DNA breaks to specific genomic regions.These breaks can then be repaired via recombination to edit, delete,insert, or otherwise modify the DNA of a host organism. In someembodiments, TALENSs are used alone for gene editing (e.g., for thedeletion or disruption of a gene). In other embodiments, TALs are usedin conjunction with donor sequences and/or other recombination factorproteins that will assist in the Non-homologous end joining (NHEJ)process to replace the targeted DNA region. For more information on theTAL-mediated gene editing compositions and methods of the presentdisclosure, see U.S. Pat. Nos. 8,440,432; 8,450,471; 8,586,526;8,586,363; 8,592,645; 8,697,853; 8,704,041; 8,921,112; and 8,912,138,each of which is hereby incorporated in its entirety for all purposes.

In some embodiments, the breeding and selection methods of the presentdisclosure are compatible with plants that have been modified throughClustered Regularly Interspaced Short Palindromic Repeats (CRISPR) orCRISPR-associated (Cas) gene editing tools. CRISPR proteins wereoriginally discovered as bacterial adaptive immunity systems whichprotected bacteria against viral and plasmid invasion. There are atleast three main CRISPR system types (Type I, II, and III) and at least10 distinct subtypes (Makarova, K. S., et. al., Nat Rev Microbiol. 2011May 9; 9(6):467-477). Type I and III systems use Cas protein complexesand short guide polynucleotide sequences to target selected DNA regions.Type II systems rely on a single protein (e.g. Cas9) and the targetingguide polynucleotide, where a portion of the 5′ end of a guide sequenceis complementary to a target nucleic acid. For more information on theCRISPR gene editing compositions and methods of the present disclosure,see U.S. Pat. Nos. 8,697,359; 8,889,418; 8,771,945; and 8,871,445, eachof which is hereby incorporated in its entirety for all purposes.

In some embodiments, the breeding and selection methods of the presentdisclosure are compatible with plants that have been modified throughmeganucleases. In some embodiments, meganucleases are engineeredendonucleases capable of targeting selected DNA sequences and inducingDNA breaks. In some embodiments, new meganucleases targeting specificregions are developed through recombinant techniques which combine theDNA binding motifs from various other identified nucleases. In otherembodiments, new meganucleases are created through semi-rationalmutational analysis, which attempts to modify the structure of existingbinding domains to obtain specificity for additional sequences. For moreinformation on the use of meganucleases for genome editing, see Silva etal., 2011 Current Gene Therapy 11 pg 11-27; and Stoddard et al., 2014Mobile DNA 5 pg 7, each of which is hereby incorporated in its entiretyfor all purposes.

Plant Transformation

Hemp plants of the present disclosure, such as ‘05.09.24.S1’ can befurther modified by introducing one or more transgenes which whenexpressed lead to desired phenotypes. The most common method for theintroduction of new genetic material into a plant genome involves theuse of living cells of the bacterial pathogen Agrobacterium tumefaciensto literally inject a piece of DNA, called transfer or T-DNA, intoindividual plant cells (usually following wounding of the tissue) whereit is targeted to the plant nucleus for chromosomal integration. Thereare numerous patents governing Agrobacterium mediated transformation andparticular DNA delivery plasmids designed specifically for use withAgrobacterium—for example, U.S. Pat. No. 4,536,475, EP0265556,EP0270822, WO8504899, WO8603516, U.S. Pat. No. 5,591,616, EP0604662,EP0672752, WO8603776, WO9209696, WO9419930, WO9967357, U.S. Pat. No.4,399,216, WO8303259, U.S. Pat. No. 5,731,179, EP068730, WO9516031, U.S.Pat. Nos. 5,693,512, 6,051,757 and EP904362A1. Agrobacterium-mediatedplant transformation involves as a first step the placement of DNAfragments cloned on plasmids into living Agrobacterium cells, which arethen subsequently used for transformation into individual plant cells.Agrobacterium-mediated plant transformation is thus an indirect planttransformation method. Methods of Agrobacterium-mediated planttransformation that involve using vectors with no T-DNA are also wellknown to those skilled in the art and can have applicability in thepresent disclosure. See, for example, U.S. Pat. No. 7,250,554, whichutilizes P-DNA instead of T-DNA in the transformation vector.

Direct plant transformation methods using DNA have also been reported.The first of these to be reported historically is electroporation, whichutilizes an electrical current applied to a solution containing plantcells (M. E. Fromm et al., Nature, 319, 791 (1986); H. Jones et al.,Plant Mol. Biol., 13, 501 (1989) and H. Yang et al., Plant Cell Reports,7, 421 (1988). Another direct method, called “biolistic bombardment”,uses ultrafine particles, usually tungsten or gold, that are coated withDNA and then sprayed onto the surface of a plant tissue with sufficientforce to cause the particles to penetrate plant cells, including thethick cell wall, membrane and nuclear envelope, but without killing atleast some of them (U.S. Pat. Nos. 5,204,253, 5,015,580). A third directmethod uses fibrous forms of metal or ceramic consisting of sharp,porous or hollow needle-like projections that literally impale thecells, and also the nuclear envelope of cells. Both silicon carbide andaluminum borate whiskers have been used for plant transformation (Mizunoet al., 2004; Petolino et al., 2000; U.S. Pat. No. 5,302,523 USApplication 20040197909) and also for bacterial and animaltransformation (Kaepler et al., 1992; Raloff, 1990; Wang, 1995). Thereare other methods reported, and undoubtedly, additional methods will bedeveloped. However, the efficiencies of each of these indirect or directmethods in introducing foreign DNA into plant cells are invariablyextremely low, making it necessary to use some method for selection ofonly those cells that have been transformed, and further, allowinggrowth and regeneration into plants of only those cells that have beentransformed.

For efficient plant transformation, a selection method must be employedsuch that whole plants are regenerated from a single transformed celland every cell of the transformed plant carries the DNA of interest.These methods can employ positive selection, whereby a foreign gene issupplied to a plant cell that allows it to utilize a substrate presentin the medium that it otherwise could not use, such as mannose or xylose(for example, refer U.S. Pat. Nos. 5,767,378; 5,994,629). Moretypically, however, negative selection is used because it is moreefficient, utilizing selective agents such as herbicides or antibioticsthat either kill or inhibit the growth of nontransformed plant cells andreducing the possibility of chimeras. Resistance genes that areeffective against negative selective agents are provided on theintroduced foreign DNA used for the plant transformation. For example,one of the most popular selective agents used is the antibiotickanamycin, together with the resistance gene neomycin phosphotransferase(nptII), which confers resistance to kanamycin and related antibiotics(see, for example, Messing & Vierra, Gene 19: 259-268 (1982); Bevan etal., Nature 304:184-187 (1983)). However, many different antibiotics andantibiotic resistance genes can be used for transformation purposes(refer U.S. Pat. Nos. 5,034,322, 6,174,724 and 6,255,560). In addition,several herbicides and herbicide resistance genes have been used fortransformation purposes, including the bar gene, which confersresistance to the herbicide phosphinothricin (White et al., Nucl AcidsRes 18: 1062 (1990), Spencer et al., Theor Appl Genet 79: 625-631(1990),U.S. Pat. Nos. 4,795,855, 5,378,824 and 6,107,549). In addition, thedhfr gene, which confers resistance to the anticancer agentmethotrexate, has been used for selection (Bourouis et al., EMBO J.2(7): 1099-1104 (1983).

Genes can be introduced in a site directed fashion using homologousrecombination. Homologous recombination permits site specificmodifications in endogenous genes and thus inherited or acquiredmutations may be corrected, and/or novel alterations may be engineeredinto the genome. Homologous recombination and site-directed integrationin plants are discussed in, for example, U.S. Pat. Nos. 5,451,513,5,501,967 and 5,527,695.

Methods of producing transgenic plants are well known to those ofordinary skill in the art. Transgenic plants can now be produced by avariety of different transformation methods including, but not limitedto, electroporation; microinjection; microprojectile bombardment, alsoknown as particle acceleration or biolistic bombardment; viral-mediatedtransformation; and Agrobacterium-mediated transformation. See, forexample, U.S. Pat. Nos. 5,405,765; 5,472,869; 5,538,877; 5,538,880;5,550,318; 5,641,664; and 5,736,369; and International PatentApplication Publication Nos. WO/2002/038779 and WO/2009/117555; Lu etal., (Plant Cell Reports, 2008, 27:273-278); Watson et al., RecombinantDNA, Scientific American Books (1992); Hinchee et al., Bio/Tech.6:915-922 (1988); McCabe et al., Bio/Tech. 6:923-926 (1988); Toriyama etal., Bio/Tech. 6:1072-1074 (1988); Fromm et al., Bio/Tech. 8:833-839(1990); Mullins et al., Bio/Tech. 8:833-839 (1990); Hiei et al., PlantMolecular Biology 35:205-218 (1997); Ishida et al., Nature Biotechnology14:745-750 (1996); Zhang et al., Molecular Biotechnology 8:223-231(1997); Ku et al., Nature Biotechnology 17:76-80 (1999); and, Raineri etal., Bio/Tech. 8:33-38 (1990)), each of which is expressly incorporatedherein by reference in their entirety. Other references teaching thetransformation of cannabis plants and the production of callus tissueinclude Raharjo et al 2006, “Callus Induction and PhytochemicalCharacterization of Cannabis sativa Cell Suspension Cultures”, Indo. J.Chem 6 (1) 70-74; and “The biotechnology of Cannabis sativa” by Sam R.Zwenger, electronically published April, 2009.

Microprojectile bombardment is also known as particle acceleration,biolistic bombardment, and the gene gun (Biolistic® Gene Gun). The genegun is used to shoot pellets that are coated with genes (e.g., fordesired traits) into plant seeds or plant tissues in order to get theplant cells to then express the new genes. The gene gun uses an actualexplosive (.22 caliber blank) to propel the material. Compressed air orsteam may also be used as the propellant. The Biolistic® Gene Gun wasinvented in 1983-1984 at Cornell University by John Sanford, EdwardWolf, and Nelson Allen. It and its registered trademark are now owned byE. I. du Pont de Nemours and Company. Most species of plants have beentransformed using this method.

Agrobacterium tumefaciens is a naturally occurring bacterium that iscapable of inserting its DNA (genetic information) into plants,resulting in a type of injury to the plant known as crown gall. Mostspecies of plants can now be transformed using this method, includingcucurbitaceous species. A transgenic plant formed using Agrobacteriumtransformation methods typically contains a single gene on onechromosome, although multiple copies are possible. Such transgenicplants can be referred to as being hemizygous for the added gene. A moreaccurate name for such a plant is an independent segregant, because eachtransformed plant represents a unique T-DNA integration event (U.S. Pat.No. 6,156,953). A transgene locus is generally characterized by thepresence and/or absence of the transgene. A heterozygous genotype inwhich one allele corresponds to the absence of the transgene is alsodesignated hemizygous (U.S. Pat. No. 6,008,437).

General transformation methods, and specific methods for transformingcertain plant species (e.g., maize) are described in U.S. Pat. Nos.4,940,838, 5,464,763, 5,149,645, 5,501,967, 6,265,638, 4,693,976,5,635,381, 5,731,179, 5,693,512, 6,162,965, 5,693,512, 5,981,840,6,420,630, 6,919,494, 6,329,571, 6,215,051, 6,369,298, 5,169,770,5,376,543, 5,416,011, 5,569,834, 5,824,877, 5,959,179, 5,563,055, and5,968,830, each of which is incorporated herein by reference in itsentirety for all purposes.

Non-limiting examples of methods for transforming cannabis plants andcannabis tissue culture methods are described in Zweger (TheBiotechnology of Cannabis sativa, April 2009); MacKinnon (Genetictransformation of Cannabis sativa Linn: a multipurpose fiber crop,doctoral thesis, University of Dundee, Scotland, 2003), MacKinnon et al.(Progress towards transformation of fiber hemp, Scottish Crop Research,2000), and US 20120311744, each of which is herein incorporated byreference in its entirety for all purposes. The transformation can bephysical, chemical and/or biological.

Herbicide Resistance

Numerous herbicide resistance genes are known and may be employed withthe invention. A non-limiting example is a gene conferring resistance toa herbicide that inhibits the growing point or meristem such asimidazolinone or sulfonylurea herbicides. As imidazolinone andsulfonylurea herbicides are acetolactate synthase (ALS)-inhibitingherbicides that prevent the formation of branched chain amino acids,exemplary genes in this category code for ALS and AHAS enzymes asdescribed, for example, by Lee et al., EMBO J., 7:1241, 1988; Gleen etal., Plant Molec. Biology, 18:1185, 1992; and Miki et al., Theor. Appl.Genet., 80:449, 1990. As a non-limiting example, a gene may be employedto confer resistance to the exemplary sulfonylurea herbicidenicosulfuron.

Resistance genes for glyphosate (resistance conferred by mutant5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyltransferase (PAT) and Streptomyces hygroscopicusphosphinothricin acetyltransferase (bar) genes) may also be used. See,for example, U.S. Pat. No. 4,940,835 to Shah et al., which discloses thenucleotide sequence of a form of EPSPS that can confer glyphosateresistance. Non-limiting examples of EPSPS transformation eventsconferring glyphosate resistance are provided by U.S. Pat. Nos.6,040,497 and 7,632,985. The MON89788 event disclosed in U.S. Pat. No.7,632,985 in particular is beneficial in conferring glyphosate tolerancein combination with an increase in average yield relative to priorevents.

A DNA molecule encoding a mutant aroA gene can be obtained under ATCCAccession No. 39256, and the nucleotide sequence of the mutant gene isdisclosed in U.S. Pat. No. 4,769,061 to Comai. A hygromycin Bphosphotransferase gene from E. coli that confers resistance toglyphosate in tobacco callus and plants is described in Penaloza-Vazquezet al., Plant Cell Reports, 14:482, 1995. European Patent ApplicationPublication No. EP0333033 to Kumada et al., and U.S. Pat. No. 4,975,374to Goodman et al., disclose nucleotide sequences of glutamine synthetasegenes that confer resistance to herbicides such as L-phosphinothricin.The nucleotide sequence of a phosphinothricin acetyltransferase gene isprovided in European Patent Application Publication No. EP0242246 toLeemans et al. DeGreef et al. (Biotechnology, 7:61, 1989) describe theproduction of transgenic plants that express chimeric bar genes codingfor phosphinothricin acetyl transferase activity. Exemplary genesconferring resistance to a phenoxy class herbicide haloxyfop and acyclohexanedione class herbicide sethoxydim are the Acct-S1, Acct-S2 andAcct-S3 genes described by Marshall et al., (Theor. Appl. Genet.,83:435, 1992). As a non-limiting example, a gene may confer resistanceto other exemplary phenoxy class herbicides that include, but are notlimited to, quizalofop-p-ethyl and 2,4-dichlorophenoxyacetic acid(2,4-D).

Genes are also known that confer resistance to herbicides that inhibitphotosynthesis such as, for example, triazine herbicides (psbA and gs+genes) and benzonitrile herbicides (nitrilase gene). As a non-limitingexample, a gene may confer resistance to the exemplary benzonitrileherbicide bromoxynil. Przibila et al. (Plant Cell, 3:169, 1991) describethe transformation of Chlamydomonas with plasmids encoding mutant psbAgenes. Nucleotide sequences for nitrilase genes are disclosed in U.S.Pat. No. 4,810,648 to Stalker, and DNA molecules containing these genesare available under ATCC Accession Nos. 53435, 67441, and 67442. Cloningand expression of DNA coding for a glutathione S-transferase isdescribed by Hayes et al. (Biochem. J., 285:173, 1992).4-hydroxyphenylpyruvate dioxygenase (HPPD) is a target of theHPPD-inhibiting herbicides, which deplete plant plastoquinone andvitamin E pools. Rippert et al. (Plant Physiol., 134:92, 2004) describesan HPPD-inhibitor resistant tobacco plant that was transformed with ayeast-derived prephenate dehydrogenase (PDH) gene. Protoporphyrinogenoxidase (PPO) is the target of the PPO-inhibitor class of herbicides; aPPO-inhibitor resistant PPO gene was recently identified in Amaranthustuberculatus (Patzoldt et al., PNAS, 103(33):12329, 2006). The herbicidemethyl viologen inhibits CO₂ assimilation. Foyer et al. (Plant Physiol.,109:1047, 1995) describe a plant overexpressing glutathione reductase(GR) that is resistant to methyl viologen treatment.

Siminszky (Phytochemistry Reviews, 5:445, 2006) describes plantcytochrome P450-mediated detoxification of multiple, chemicallyunrelated classes of herbicides. Modified bacterial genes have beensuccessfully demonstrated to confer resistance to atrazine, a herbicidethat binds to the plastoquinone-binding membrane protein Q_(B) inphotosystem II to inhibit electron transport. See, for example, studiesby Cheung et al. (PNAS, 85:391, 1988), describing tobacco plantsexpressing the chloroplast psbA gene from an atrazine-resistant biotypeof Amaranthus hybridus fused to the regulatory sequences of a nucleargene, and Wang et al. (Plant Biotech. J., 3:475, 2005), describingtransgenic alfalfa, Arabidopsis, and tobacco plants expressing the atzAgene from Pseudomonas sp. that were able to detoxify atrazine.

Bayley et al. (Theor. Appl. Genet., 83:645, 1992) describe the creationof 2,4-D-resistant transgenic tobacco and cotton plants using the 2,4-Dmonooxygenase gene tfdA from Alcaligenes eutrophus plasmid pJP5. U.S.Patent Application Publication No. 20030135879 describes the isolationof a gene for dicamba monooxygenase (DMO) from Psueodmonas maltophiliathat is involved in the conversion of dicamba to a non-toxic3,6-dichlorosalicylic acid and thus may be used for producing plantstolerant to this herbicide.

Other examples of herbicide resistance have been described, forinstance, in U.S. Pat. Nos. 6,803,501; 6,448,476; 6,248,876; 6,225,114;6,107,549; 5,866,775; 5,804,425; 5,633,435; 5,463,175.

Disease and Pest Resistance

Plant defenses are often activated by specific interaction between theproduct of a disease resistance gene (R) in the plant and the product ofa corresponding avirulence (Avr) gene in the pathogen. A plant line canbe transformed with a cloned resistance gene to engineer plants that areresistant to specific pathogen strains. See, for example Jones et al.(Science, 266:789-793, 1994) (cloning of the tomato Cf-9 gene forresistance to Cladosporium flavum); Martin et al. (Science,262:1432-1436, 1993) (tomato Pto gene for resistance to Pseudomonassyringae pv. tomato); and Mindrinos et al. (Cell, 78(6):1089-1099, 1994)(Arabidopsis RPS2 gene for resistance to Pseudomonas syringae).

A viral-invasive protein or a complex toxin derived therefrom may alsobe used for viral disease resistance. For example, the accumulation ofviral coat proteins in transformed plant cells imparts resistance toviral infection and/or disease development effected by the virus fromwhich the coat protein gene is derived and related viruses. See Beachyet al. (Ann. Rev. Phytopathol., 28:451, 1990). Coat protein-mediatedresistance has been conferred upon transformed plants against alfalfamosaic virus, cucumber mosaic virus, tobacco streak virus, potato virusX, potato virus Y, tobacco etch virus, tobacco rattle virus, and tobaccomosaic virus.

A virus-specific antibody may also be used. See, for example,Tavladoraki et al. (Nature, 366:469-472, 1993), who show that transgenicplants expressing recombinant antibody genes are protected from virusattack. Virus resistance has also been described in, for example, U.S.Pat. Nos. 6,617,496; 6,608,241; 6,015,940; 6,013,864; 5,850,023 and5,304,730. Additional means of inducing whole-plant resistance to apathogen include modulation of the systemic acquired resistance (SAR) orpathogenesis related (PR) genes, for example genes homologous to theArabidopsis thaliana NIM1/NPR1/SAI1, and/or by increasing salicylic acidproduction (Ryals et al., Plant Cell, 8:1809-1819, 1996).

Logemann et al. (Biotechnology, 10:305-308, 1992), for example, disclosetransgenic plants expressing a barley ribosome-inactivating gene thathave an increased resistance to fungal disease. Plant defensins may beused to provide resistance to fungal pathogens (Thomma et al., Planta,216:193-202, 2002). Other examples of fungal disease resistance areprovided in U.S. Pat. Nos. 6,653,280; 6,573,361; 6,506,962; 6,316,407;6,215,048; 5,516,671; 5,773,696; 6,121,436; and 6,316,407.

Nematode resistance has been described in, for example, U.S. Pat. No.6,228,992, and bacterial disease resistance has been described in, forexample, U.S. Pat. No. 5,516,671.

The use of the herbicide glyphosate for disease control in hemp plantscontaining event MON89788, which confers glyphosate tolerance, has alsobeen described in U.S. Pat. No. 7,608,761.

Insect Resistance

One example of an insect resistance gene includes a Bacillusthuringiensis protein, a derivative thereof, or a synthetic polypeptidemodeled thereon. See, for example, Geiser et al. (Gene, 48(1):109-118,1986), who disclose the cloning and nucleotide sequence of a Bacillusthuringiensis δ-endotoxin gene. Moreover, DNA molecules encodingδ-endotoxin genes can be purchased from the American Type CultureCollection, Manassas, Va., for example, under ATCC Accession Nos. 40098,67136, 31995 and 31998. Another example is a lectin. See, for example,Van Damme et al., (Plant Molec. Biol., 24:825-830, 1994), who disclosethe nucleotide sequences of several Clivia miniata mannose-bindinglectin genes. A vitamin-binding protein may also be used, such as, forexample, avidin. See PCT Application No. US93/06487, the contents ofwhich are hereby incorporated by reference. This application teaches theuse of avidin and avidin homologues as larvicides against insect pests.

Yet another insect resistance gene is an enzyme inhibitor, for example,protease, proteinase, or amylase inhibitors. See, for example, Abe etal. (J. Biol. Chem., 262:16793-16797, 1987) describing the nucleotidesequence of a rice cysteine proteinase inhibitor; Linthorst et al.(Plant Molec. Biol., 21:985-992, 1993) describing the nucleotidesequence of a cDNA encoding tobacco proteinase inhibitor I; and Sumitaniet al. (Biosci. Biotech. Biochem., 57:1243-1248, 1993) describing thenucleotide sequence of a Streptomyces nitrosporeus α-amylase inhibitor.

An insect-specific hormone or pheromone may also be used. See, forexample, the disclosure by Hammock et al. (Nature, 344:458-461, 1990) ofbaculovirus expression of cloned juvenile hormone esterase, aninactivator of juvenile hormone; Gade and Goldsworthy (Eds.Physiological System in Insects, Elsevier Academic Press, Burlington,Mass., 2007), describing allostatins and their potential use in pestcontrol; and Palli et al. (Vitam. Horm., 73:59-100, 2005), disclosinguse of ecdysteroid and ecdysteroid receptor in agriculture. The diuretichormone receptor (DHR) was identified in Price et al. (Insect Mol.Biol., 13:469-480, 2004) as another potential candidate target ofinsecticides.

Still other examples include an insect-specific antibody or animmunotoxin derived therefrom and a developmental-arrestive protein. SeeTaylor et al. (Seventh Int'l Symposium on Molecular Plant-MicrobeInteractions, Edinburgh, Scotland, Abstract W97, 1994), who describedenzymatic inactivation in transgenic tobacco via production ofsingle-chain antibody fragments. Numerous other examples of insectresistance have been described. See, for example, U.S. Pat. Nos.6,809,078; 6,713,063; 6,686,452; 6,657,046; 6,645,497; 6,642,030;6,639,054; 6,620,988; 6,593,293; 6,555,655; 6,538,109; 6,537,756;6,521,442; 6,501,009; 6,468,523; 6,326,351; 6,313,378; 6,284,949;6,281,016; 6,248,536; 6,242,241; 6,221,649; 6,177,615; 6,156,573;6,153,814; 6,110,464; 6,093,695; 6,063,756; 6,063,597; 6,023,013;5,959,091; 5,942,664; 5,942,658, 5,880,275; 5,763,245 and 5,763,241.

Resistance to Abiotic Stress

Abiotic stress includes dehydration or other osmotic stress, salinity,high or low light intensity, high or low temperatures, submergence,exposure to heavy metals, and oxidative stress.Delta-pyrroline-5-carboxylate synthetase (P5CS) from mothbean has beenused to provide protection against general osmotic stress.Mannitol-1-phosphate dehydrogenase (mt1D) from E. coli has been used toprovide protection against drought and salinity. Choline oxidase (codAfrom Arthrobactor globiformis) can protect against cold and salt. E.coli choline dehydrogenase (betA) provides protection against salt.Additional protection from cold can be provided by omega-3-fatty aciddesaturase (fad7) from Arabidopsis thaliana. Trehalose-6-phosphatesynthase and levan sucrase (SacB) from yeast and Bacillus subtilis,respectively, can provide protection against drought (summarized fromAnnex II Genetic Engineering for Abiotic Stress Tolerance in Plants,Consultative Group On International Agricultural Research TechnicalAdvisory Committee). Overexpression of superoxide dismutase can be usedto protect against superoxides, see U.S. Pat. No. 5,538,878.

Additional Traits

Additional traits can be introduced into the hemp variety of the presentinvention. A non-limiting example of such a trait is a coding sequencewhich decreases RNA and/or protein levels. The decreased RNA and/orprotein levels may be achieved through RNAi methods, such as thosedescribed in U.S. Pat. No. 6,506,559.

Another trait that may find use with the hemp variety of the inventionis a sequence which allows for site-specific recombination. Examples ofsuch sequences include the FRT sequence used with the FLP recombinase(Zhu and Sadowski, J. Biol. Chem., 270:23044-23054, 1995) and the LOXsequence used with CRE recombinase (Sauer, Mol. Cell. Biol.,7:2087-2096, 1987). The recombinase genes can be encoded at any locationwithin the genome of the hemp plant and are active in the hemizygousstate.

In certain embodiments hemp plants may be made more tolerant to or moreeasily transformed with Agrobacterium tumefaciens. For example,expression of p53 and iap, two baculovirus cell-death suppressor genes,inhibited tissue necrosis and DNA cleavage. Additional targets mayinclude plant-encoded proteins that interact with the Agrobacterium Virgenes; enzymes involved in plant cell wall formation; and histones,histone acetyltransferases and histone deacetylases (reviewed in Gelvin,Microbiology &Mol. Biol. Reviews, 67:16-37, 2003).

In addition to the modification of oil, fatty acid, or phytate contentdescribed above, certain embodiments may modify the amounts or levels ofother compounds. For example, the amount or composition of antioxidantscan be altered. See, for example, U.S. Pat. Nos. 6,787,618 and 7,154,029and International Patent Application Publication No. WO 00/68393, whichdisclose the manipulation of antioxidant levels, and InternationalPatent Application Publication No. WO 03/082899, which discloses themanipulation of an antioxidant biosynthetic pathway.

Additionally, seed amino acid content may be manipulated. U.S. Pat. No.5,850,016 and International Patent Application Publication No. WO99/40209 disclose the alteration of the amino acid compositions ofseeds. U.S. Pat. Nos. 6,080,913 and 6,127,600 disclose methods ofincreasing accumulation of essential amino acids in seeds.

U.S. Pat. No. 5,559,223 describes synthetic storage proteins of whichthe levels of essential amino acids can be manipulated. InternationalPatent Application Publication No. WO 99/29882 discloses methods foraltering amino acid content of proteins. International PatentApplication Publication No. WO 98/20133 describes proteins with enhancedlevels of essential amino acids. International Patent ApplicationPublication No. WO 98/56935 and U.S. Pat. Nos. 6,346,403; 6,441,274; and6,664,445 disclose plant amino acid biosynthetic enzymes. InternationalPatent Application Publication No. WO 98/45458 describes synthetic seedproteins having a higher percentage of essential amino acids thanwild-type.

U.S. Pat. No. 5,633,436 discloses plants comprising a higher content ofsulfur-containing amino acids; U.S. Pat. No. 5,885,801 discloses plantscomprising a high threonine content; U.S. Pat. Nos. 5,885,802 and5,912,414 disclose plants comprising a high methionine content; U.S.Pat. No. 5,990,389 discloses plants comprising a high lysine content;U.S. Pat. No. 6,459,019 discloses plants comprising an increased lysineand threonine content; International Patent Application Publication No.WO 98/42831 discloses plants comprising a high lysine content;International Patent Application Publication No. WO 96/01905 disclosesplants comprising a high threonine content; and International PatentApplication Publication No. WO 95/15392 discloses plants comprising ahigh lysine content.

Cannabis Hemp Extracts and Compositions

In some embodiments, the present disclosure provides for extracts andcompositions from the hemp plants of the present disclosure. Cannabisextracts or products or the present disclosure include:

Solvent reduced oils—also sometimes known as oil, BHO, CO₂ extract,among other names. This type of extract is made by soaking plantmaterial in a chemical solvent capable of solubilizing one or morechemical constituents of the plant (e.g., cannabinoids and/or terpenes).After separating the solvent from plant material, the solvent can beboiled or evaporated off, leaving the extract “oil” behind. Butane HashOil is produced by passing butane over cannabis and then letting thebutane evaporate. Rick Simpson Oil is produced through isopropyl, orethanol extraction of cannabis. The resulting substance is a wax likegolden brown paste. Another common extraction solvent for creatingcannabis oil is CO₂. Persons having skill in the art will be familiarwith CO₂ extraction techniques and devices, including those disclosed inUS 20160279183, US 2015/01505455, U.S. Pat. No. 9,730,911, and US2018/0000857.

Heat extractions—The present disclosure also teaches extracts producedvia heat-based extraction methods, such as those disclosed in US PatentApplication Nos. US 2018/0078874, US 2019/0151771, US 2019/0076753, andU.S. Pat. No. 10,159,908, each of which is hereby incorporated byreference for all purposes. In some embodiments, the plants of thepresent disclosure can be extracted by exposing tissue to a hot air gasstream that volatizes cannabinoids and/or other secondary metabolites ofthe plant, which are then condensed and recovered in tanks.

In some embodiments, the present disclosure teaches exposing plants,plant parts or plant cells to vaporizing heat. As used herein, the term“vaporizing heat” refers to heat sufficient to volatize one or moreterpene on cannabinoid components of said plant, plant part or plantcell. The boiling points for each of the cannabinoid and terpeneconstituents of a hemp plant are well known or readily ascertainable. Insome embodiments, vaporizing heat comprises 150° F., 155° F., 160° F.,165° F., 170° F., 175° F., 180° F., 185° F., 190° F., 195° F., 200° F.,205° F., 210° F., 215° F., 220° F., 225° F., 230° F., 235° F., 240° F.,245° F., 250° F., 255° F., 260° F., 265° F., 270° F., 275° F., 280° F.,285° F., 290° F., 295° F., 300° F. ° F., 305° F. ° F., 310° F. ° F.,315° F. ° F., 320° F. ° F., 325° F. ° F., 330° F. ° F., 335° F. ° F.,340° F. ° F., 345° F., or 350° F., and all ranges and subrangestherebetween.

Tinctures—are alcoholic extracts of cannabis. These are usually made bymixing cannabis material with high proof ethanol and separating outplant material. Within the dietary supplement industry “tincture” mayalso describe an oil dilution of hemp extract.

In some embodiments, the specialty cannabis of the present disclosure isextracted via methods that preserve the cannabinoid and terpenes. Inother embodiments, said methods can be used with any cannabis plants.The extracts of the present disclosure are designed to produce productsfor human or animal consumption via inhalation (via combustion,vaporization and nebulization), buccal absorption within the mouth, oraladministration (e.g., eating/drinking), and topical application deliverymethods.

The chemical extraction of specialty cannabis can be accomplishedemploying polar and non-polar solvents in various phases at varyingpressures and temperatures to selectively or comprehensively extractterpenes, cannabinoids and other compounds of flavor, fragrance orpharmacological value for use individually or combination in theformulation of our products. The solvents employed for selectiveextraction of our cultivars may include water, carbon dioxide,1,1,1,2-tetrafluoroethane, butane, propane, ethanol, isopropyl alcohol,hexane, and limonene, in combination or series. It is also possible toextract compounds of interest mechanically by sieving the plant partsthat produce those compounds. Measuring the plant part, i.e. trichomegland head, to be sieved via optical or electron microscopy can aid theselection of the optimal sieve pore size, ranging from 30 to 130microns, to capture the plant part of interest. The chemical andmechanical extraction methods of the present disclosure can be used toproduce products that combine chemical extractions with plant partscontaining compounds of interest.

The extracts of the present disclosure may also be combined with purecompounds of interest to the extractions, e.g. cannabinoids or terpenesto further enhance or modify the resulting formulation's fragrance,flavor or pharmacology. Thus, in some embodiments, the presentdisclosure teaches compositions comprising at least one ingredientextracted from the ‘05.09.24.S1’ plant. In some embodiments, extractsfrom the hemp lines of the present disclosure are combined with one ormore additional compounds. In some embodiments, extracts of the presentdisclosure, such as whole hemp extracts, or a purified cannabinoid fromsaid hemp plant, can be combined with another cannabinoid or terpene toproduce a composition.

The compositions of the present disclosure encompass many forms. In someembodiments, the present disclosure provides CBD oils and tinctures. Insome embodiments, the present disclosure provides CBD capsules. In someembodiments, the present disclosure provides CBD infused edibles, suchas gummies, gum, lollipops, taffy, cookies, brownies, ice cream,chocolate, jerky, animal dry and wet foods, animal treats, etc. In someembodiments, the present disclosure provides for cosmetics comprisingCBD, such as lip stick, balms, creams, shampoo, conditioners, lotions,rubbing oils, lubricants, Etc. In some embodiments, the CBD oilscomprise extracts from ‘05.09.24.S1’, such as solvent extracted oils,heat extracted oils. In some embodiments, the capsules comprise extractsfrom ‘05.09.24.S1’.

In some embodiments, the present disclosure teaches hemp commodityproducts, including processed hemp inflorescences, fiber, hemp extract,cannabinoids, and terpenes. As used herein, the term “processed hempinflorescences” means inflorescences from a hemp plant that have beenharvested and dried to a moisture content of less than 20% wt/wt. Insome embodiments, the processed hemp inflorescences are ground or brokenup in smaller pieces.

In some embodiments, the present disclosure teaches methods of dryinghemp inflorescence tissue. In some embodiments, inflorescences of the‘05.09.24.S1’ variety are allowed to dry in a low moisture room orcontainer. In some embodiments the room or container used to dry theinflorescences has a humidity between 45-55%. In some embodiments, thedrying occurs in the dark or low-light conditions.

In some embodiments, harvested inflorescences are hung over aclothesline, clothes hanger, clothespins, or other device capable ofsuspending the inflorescence by the stem. In some embodiments, the hunginflorescences have been partially trimmed to remove unwanted leaves. Insome embodiments, the inflorescences are arranged such that theinflorescences are not in contact with anything (to prevent rubbing offof trichomes).

In some embodiments, the drying step comprises curing. In someembodiments, curing occurs by placing trimmed inflorescences into anairtight container, sealing the containers, and placing them in a cool,dark place to continue drying.

In some embodiments, the inflorescences are dried to a 10-20% wt/wtmoisture content. Persons having skill in the art will be familiar withmethods for measuring moisture content of plant material. In someembodiments, moisture content can be measured using an FourrierTransformation Infrared spectroscopy. In some embodiments, moisturecontent can be measured by weighing a sample, and placing the sample ina sealed container with a water-absorbing material, and then measuringthe weight of the completely dried inflorescences a few days later. Insome embodiments, the moisture content of flower material can bedetermined by measuring capacitance.

DEPOSIT INFORMATION

A deposit of the ‘05.09.24.S1’ hemp cultivar is maintained by BiotechInstitute, LLC, 5655 Lindero Canyon Road, Suite 226, Westlake Village,Calif. 91362, USA. In addition, a sample of 625 seeds of the‘05.09.24.S1’ variety of this disclosure has been deposited with anInternational Depositary Authority as established under the BudapestTreaty according to 37 CFR 1.803(a)(1). Applicant has deposited seeds atthe Provasoli-Guillard National Center for Marine Algae and Microbiota(NCMA), located at the Bigelow Laboratory for Ocean Science at 60Bigelow Drive East Boothbay, Me. 04544.

The ‘05.09.24.S1’ seeds have been deposited under the Budapest Treaty asNCMA No. 202202006 on Feb. 3, 2022.

To satisfy the enablement requirements of 35 U.S.C. 112, and to certifythat the deposit of the isolated strain (i.e., hemp plant) of thepresent disclosure meets the criteria set forth in 37 C.F.R. 1.801-1.809and Manual of Patent Examining Procedure (MPEP) 2402-2411.05, Applicantshereby make the following statements regarding the deposited‘05.09.24.S1’ hemp cultivar (deposited as NCMA No. 202202006):

-   -   1. During the pendency of this application, access to the        disclosure will be afforded to the Commissioner upon request;    -   2. All restrictions on availability to the public will be        irrevocably removed upon granting of the patent under conditions        specified in 37 CFR 1.808;    -   3. The deposit will be maintained in a public repository for a        period of 30 years or 5 years after the last request or for the        effective life of the patent, whichever is longer;    -   4. A test of the viability of the biological material at the        time of deposit will be conducted by the public depository under        37 C.F.R. 1.807; and    -   5. The deposit will be replaced if it should ever become        unavailable.

Access to this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved by affording access to a deposit of at least 625 seeds of thesame variety with the NCMA.

Unless defined otherwise, all technical and scientific terms herein havethe same meaning as commonly understood by one of ordinary skill in theart to which this invention belongs. Although any methods and materials,similar or equivalent to those described herein, can be used in thepractice or testing of the present invention, the non-limiting exemplarymethods and materials are described herein.

All publications and patent applications mentioned in the specificationare indicative of the level of those skilled in the art to which thisinvention pertains. All publications and patent applications are hereinincorporated by reference to the same extent as if each individualpublication or patent application was specifically and individuallyindicated to be incorporated by reference. Nothing herein is to beconstrued as an admission that the present disclosure is not entitled toantedate such publication by virtue of prior disclosure.

Many modifications and other embodiments of the disclosures set forthherein will come to mind to one skilled in the art to which thesedisclosures pertain having the benefit of the teachings presented in theforegoing descriptions and the associated drawings. Therefore, it is tobe understood that the disclosures are not to be limited to the specificembodiments disclosed and that modifications and other embodiments areintended to be included within the scope of the appended claims.Although specific terms are employed herein, they are used in a genericand descriptive sense only and not for purposes of limitation.

While the disclosure has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations of the disclosure following, in general, theprinciples of the disclosure and including such departures from thepresent disclosure as come within known or customary practice within theart to which the disclosure pertains and as may be applied to theessential features hereinbefore set forth and as follows in the scope ofthe appended claims.

Example 1. Cannabinoid Potency Tests

Samples of inflorescences from the ‘05.09.24.S1’ hemp line harvested atmaturity were provided for cannabinoid potency analysis. One hundredseventy eight samples of the ‘05.09.24.S1’ inflorescences were analyzedusing high performance liquid chromatography. The results of theseanalyses are provided below in Table 6.

These analyses were conducted in bulk to establish the variety'scompliance with hemp regulations (i.e., that the plants accumulated lessthan 0.3% THC). The analyzed tissues were taken from various parts ofthe plant, and without any trimming to remove non-cannabinoid producingtissue. Thus, the cannabinoid and terpene analyses of Examples 1 and 2are not reflective of the uniformity of cannabinoid and terpeneaccumulation.

TABLE 6 Cannabinoid Potency Analysis Results for ‘05.09.24.S1’ Analysis‘05.09.24.S1’ BIHEMP 050924 THC Potential About 0.0% About 0.21% toabout 0.26% to about 0.43% CBD Potential Median of about 4.39% About5.02% to about 10.86%

Example 2. Terpene Profile Tests

Samples of inflorescences from the ‘05.09.24.S1’ hemp line harvested atmaturity were provided for terpene profile analysis. One hundred seventyeight samples of the ‘05.09.24.S1’ inflorescences were analyzed usinggas chromatography. The results of these analyses are provided below inTable 7.

TABLE 7 Terpene Profile Analysis Results for ‘05.09.24.S1’ 05.09.24.S1BIHEMP 050924 Compound % (w/w) % (w/w) thujene 0 0 alpha-pinene   0 to0.03 0.01 to 0.04 camphene   0 to 0.01   0 to 0.01 sabinene 0 0beta-pinene   0 to 0.05 0.01 to 0.07 myrcene   0 to 0.56   0 to 0.12ethyl caproate 0 0 alpha-phellandrene 0 0 carene 0 0 hexyl acetate 0 0alpha-terpinene 0 0 cymene 0 0 limonene   0 to 0.36 0.06 to 0.62beta-phellandrene 0 0 cineole   0 to 0.04 0 cis-ocimene 0 0trans-ocimene 0   0 to 0.12 gamma-terpinene 0 0 linalool oxide 0 0terpinolene 0   0 to 0.02 fenchone 0 0 linalool   0 to 0.18 0.02 to 0.16hexyl propanoate 0 0 fenchol   0 to 0.03 0.01 to 0.06 MT_1124   0 to0.02 0 isoborneol 0 0 (-)borneol 0   0 to 0.02 hexyl butyrate 0 0alpha-terpineol   0 to 0.03 0.01 to 0.06 ethyl-octanoate 0 0 citronellol0 0 hexyl hexanoate 0   0 to 0.03 octyl butyrate   0 to 0.02 0 betacaryophyllene 0.11 to 0.93 0.12 to 0.60 gamma-elemene 0 0 bergamotene  0 to 0.18 n/a alpha-guaiene   0 to 0.22 n/a beta-farnesene   0 to 0.35n/a alpha-humulene 0.04 to 0.38 0.03 to 0.38 caryophyllene oxide   0 to0.04   0 to 0.03 alpha-bisabolol   0 to 0.07 0.01 to 0.03

INCORPORATION BY REFERENCE

All references, articles, publications, patents, patent publications,and patent applications cited herein are incorporated by reference intheir entireties for all purposes. However, mention of any reference,article, publication, patent, patent publication, and patent applicationcited herein is not, and should not be taken as, an acknowledgment orany form of suggestion that they constitute valid prior art or form partof the common general knowledge in any country in the world.

Numbered Embodiments

Further embodiments contemplated by the disclosure are listed below.

1. A seed, plant, plant part, or plant cell of hemp plant varietydesignated ‘05.09.24.S1’, wherein seed of the variety has been depositedunder NCMA No. 202202006.

2. The hemp plant part of embodiment 1, wherein the plant part is aninflorescence.

3. A hemp plant or a plant part or a plant cell thereof, having all ofthe characteristics of the hemp plant variety designated ‘05.09.24.S1’deposited under NCMA No. 202202006.

4. A hemp plant, or a plant part or a plant cell thereof, having all ofthe physiological and morphological characteristics of the hemp plant ofany one of embodiments 1-3.

5. A hemp plant, or a part or a plant cell thereof, having all of thephysiological and morphological characteristics of the hemp plantvariety designated ‘05.09.24. S1’, wherein seed of said variety wasdeposited under NCMA No. 202202006.

6. A tissue culture of regenerable cells produced from the plant, plantpart or plant cell of any one of embodiments 1-5, wherein a new plantregenerated from the tissue culture has all of the characteristics ofthe hemp plant variety designated ‘05.09.24.S1’ deposited under NCMA No.202202006 when grown under the same environmental conditions.

7. A hemp plant regenerated from the tissue culture of embodiment 6,said plant having all the characteristics of the hemp plant varietydesignated ‘05.09.24.S1’ deposited under NCMA No. 202202006 when grownunder the same environmental conditions.

8. A hemp plant regenerated from the tissue culture of embodiment 6,wherein the regenerated plant has all of the characteristics of the hempplant variety designated ‘05.09.24.S1’, wherein seed of said variety wasdeposited under NCMA No. 202202006.

9. A method for producing a hemp seed, comprising a) selfing the hempplant of any one of embodiments 1-5 and 7-8, and b) harvesting theresultant hemp seed.

10. A hemp seed produced by the method of embodiment 9.

11. A method for producing a hemp seed comprising crossing the hempplant of any one of embodiments 1-5 and 7-8 with a second, distinctplant.

12. An F1 hemp seed produced by the method of embodiment 11.

13. An F1 hemp plant, or a part or a plant cell thereof, produced bygrowing the seed of embodiment 12.

14. A method of producing a hemp plant derived from the variety‘05.09.24.S1’ comprising: a) crossing the plant of any one ofembodiments 1-5 and 7-8, with itself or a second plant to produceprogeny seed; b) growing the progeny seed to produce a progeny plant andcrossing the progeny plant with itself or a second plant to producefurther progeny seed; and c) repeating steps (a) and (b) with sufficientinbreeding until a seed of an hemp plant derived from the variety‘05.09.24.S1’ is produced.

15. The method of embodiment 14, further comprising crossing the hempplant derived from the variety ‘05.09.24.S1,’ with a plant of adifferent genotype to produce seed of a hybrid plant derived from thehemp variety ‘05.09.24.S1’

16. A method for producing nucleic acids, the method comprisingisolating nucleic acids from the seed, plant, plant part, or plant cellof any one of embodiments 1-15.

17. The hemp plant of any one of embodiments 1-5 and 7-8, comprising asingle locus conversion and otherwise all of the characteristics of thehemp plant of any one of embodiments 1-5 and 7-8 when grown in the sameenvironmental conditions.

18. The hemp plant of embodiment 17, wherein the single locus conversionconfers said plant with herbicide resistance.

19. The hemp plant of embodiment 17, wherein the single locus conversionis an artificially mutated gene or nucleotide sequence.

20. The hemp plant of embodiment 17, wherein the single locus conversionis a gene that has been modified through the use of breeding techniques.

21. A cultivar of hemp designated ‘05.09.24.S1’ as described anddetailed herein.

22. A method of producing a cannabinoid extract, said method comprisingthe steps a) contacting the plant of any one of embodiments 1-5 and 7-8with a solvent, thereby producing a cannabinoid extract.

23. A dry, sinsemilla non-viable plant or part thereof, wherein seed ofhemp plants producing said dry plant and part thereof has been depositedunder NCMA No. 202202006.

24. An assemblage of dry, non-viable sinsemilla female inflorescencesfrom a hemp plant variety designated ‘05.09.24. S1’ wherein seed of thevariety has been deposited under NCMA No. 202202006.

25. The dry, non-viable plant part of embodiment 23 or 24, wherein theplant part is an inflorescence.

26. The dry, non-viable plant part of embodiment 23 or 24, wherein theplant part is a trichome.

27. Dry, non-viable kief powder comprising cannabidiol (CBD), whereinseed of hemp plants producing said kief has been deposited under NCMANo. 202202006.

28. A method of producing a hemp plant with cannabidiol (CBD), saidmethod comprising propagating a vegetative cutting from a hemp plantvariety designated ‘05.09.24.S1’ wherein seed of the variety has beendeposited under NCMA No. 202202006.

29. The hemp plant with CBD, produced according to the methods ofembodiment 28.

30. The hemp plant of embodiment 5, wherein the plant is asexuallyreproduced.

31. A method for producing a hemp plant with inflorescences that producecannabidiol (CBD), said method comprising:

propagating a vegetative cutting from a stock hemp plant, therebyproducing the hemp plant having CBD;

wherein the stock hemp plant is a product of applying a plant breedingtechnique to a variety designated ‘05.09.24.S1’, wherein seed of thevariety has been deposited under NCMA No. 202202006.

32. The method of embodiment 31, wherein said plant breeding techniqueis recurrent selection.

33. The method of embodiment 31, wherein said plant breeding techniqueis mass selection.

34. The method of embodiment 31, wherein said plant breeding techniqueis hybridization.

35. The method of embodiment 31, wherein said plant breeding techniqueis open-pollination.

36. The method of embodiment 31, wherein said plant breeding techniqueis backcrossing.

37. The method of embodiment 31, wherein said plant breeding techniqueis pedigree breeding.

38. The method of embodiment 31, wherein said plant breeding techniqueis mutation breeding, and wherein said mutation selected is spontaneousor artificially induced.

39. A method for producing a Cannabis plant derived from a hemp varietydesignated ‘05.09.24.S1’, said method comprising crossing ‘05.09.24.S1’with another Cannabis plant, thereby producing a progeny Cannabis plant;wherein seed of the ‘05.09.24.S1’ variety has been deposited under NCMANo. 202202006.

40. The method of embodiment 39, further comprising the steps of:

a) crossing the progeny Cannabis plant from a previous step with itselfor another Cannabis plant to produce a progeny Cannabis plant of asubsequent generation;

b) repeating step (a) for one or more additional generations to producea Cannabis plant further derived from the hemp variety designated‘05.09.24.S1’.

41. The method of embodiment 39 or 40, further comprising the step ofcontacting the Cannabis plant further derived from the hemp varietydesignated ‘05.09.24.S1’ or a plant part derived therefrom with asolvent, or exposing said Cannabis plant or plant part to vaporizingheat, thereby producing a cannabinoid extract.

42. A method for producing a Cannabis plant derived from a hemp varietydesignated ‘05.09.24. S1’, said method comprising:

propagating a vegetative cutting from a stock Cannabis plant, therebyproducing the Cannabis plant derived from the hemp variety designated‘05.09.24.S1’;

wherein the stock Cannabis plant is a product of applying a plantbreeding technique to ‘05.09.24.S1’, wherein seed of the ‘05.09.24.S1’variety has been deposited under NCMA No. 202202006.

43. The method of embodiment 42, further comprising the step ofcontacting the Cannabis plant derived from the hemp variety designated‘05.09.24.S1’ or a plant part derived therefrom with a solvent, orexposing said Cannabis plant or plant part to vaporizing heat, therebyproducing a cannabinoid extract.

44. The method of embodiment 42, wherein said plant breeding techniqueis recurrent selection.

45. The method of embodiment 42, wherein said plant breeding techniqueis mass selection.

46. The method of embodiment 42, wherein said plant breeding techniqueis hybridization.

47. The method of embodiment 42, wherein said plant breeding techniqueis open-pollination.

48. The method of embodiment 42, wherein said plant breeding techniqueis backcrossing.

49. The method of embodiment 42, wherein said plant breeding techniqueis pedigree breeding.

50. The method of embodiment 42, wherein said plant breeding techniqueis mutation breeding, and wherein said mutation selected is spontaneousor artificially induced.

51. The method of embodiment 42, wherein said plant breeding techniqueis marker enhanced selection.

52. A method for producing a Cannabis plant derived from a hemp varietydesignated ‘05.09.24. S1’, said method comprising:

crossing a stock Cannabis plant with itself or another Cannabis plant,thereby producing the Cannabis plant derived from the hemp varietydesignated ‘05.09.24.S1’;

wherein the stock Cannabis plant is a product of applying a plantbreeding technique to ‘05.09.24.S1’, wherein seed of the ‘05.09.24.S1’variety has been deposited under NCMA No. 202202006.

53. The method of embodiment 52, further comprising the step ofcontacting the Cannabis plant derived from the hemp variety designated‘05.09.24.S1’ or a plant part derived therefrom with a solvent, orexposing said Cannabis plant or plant part to vaporizing heat, therebyproducing a cannabinoid extract.

54. The method of embodiment 52, wherein said plant breeding techniqueis recurrent selection.

55. The method of embodiment 52, wherein said plant breeding techniqueis mass selection.

56. The method of embodiment 52, wherein said plant breeding techniqueis hybridization.

57. The method of embodiment 52, wherein said plant breeding techniqueis open-pollination.

58. The method of embodiment 52, wherein said plant breeding techniqueis backcrossing.

59. The method of embodiment 52, wherein said plant breeding techniqueis pedigree breeding.

60. The method of embodiment 52, wherein said plant breeding techniqueis mutation breeding, and wherein said mutation selected is spontaneousor artificially induced.

61. The method of embodiment 52, wherein said plant breeding techniqueis marker enhanced selection.

62. The hemp plant of embodiment 5, wherein the plant is capable ofproducing an asexual clone of said hemp plant.

63. The hemp plant of embodiment 62, wherein the asexual clone iscapable of producing said hemp plant of embodiment 1.

64. A method of producing a commodity plant product, the methodcomprising producing the commodity plant product from the plant ofembodiment 1.

65. A method of producing a commodity plant product comprisingcollecting the commodity plant product from a seed, plant, plant part,or plant cell of hemp plant variety designated ‘05.09.24.S1’, whereinseed of the variety has been deposited under NCMA No. 202202006.

66. The method of embodiments 64 or 65, wherein the commodity plantproduct is selected from a group consisting of processed hempinflorescence, hemp fiber, hemp oil extract, terpenes, and cannabinoids.

What is claimed is:
 1. A seed, plant, plant part, or plant cell of ahemp plant variety designated ‘05.09.24.S1’, wherein seed of the varietyhas been deposited under NCMA No.
 202202006. 2. The plant part of claim1, wherein the plant part is selected from the group consisting of aleaf, a stem, an inflorescence, and a trichome.
 3. The plant part ofclaim 1, wherein the plant part is an inflorescence.
 4. A tissue cultureof regenerable cells produced from the plant, plant part or plant cellof claim
 1. 5. A hemp plant regenerated from the tissue culture of claim4, said plant having all the morphological and physiologicalcharacteristics of the hemp plant variety designated ‘05.09.24.S1’deposited under NCMA No. 202202006, when grown under the sameenvironmental conditions.
 6. A method for producing nucleic acids, themethod comprising isolating nucleic acids from the seed, plant, plantpart, or plant cell of claim
 1. 7. A plant, plant part, or plant cellcomprising a single locus conversion and otherwise all of themorphological and physiological characteristics of the hemp plantvariety designated ‘05.09.24.S1’ deposited under NCMA No. 202202006,when grown under the same environmental conditions.
 8. The plant, plantpart, or plant cell of claim 7, wherein the single locus conversioncomprises a transgene.
 9. A seed that produces the plant of claim
 8. 10.The plant, plant part, or plant cell of claim 7, wherein the singlelocus conversion confers a trait selected from the group consisting ofmale sterility, herbicide tolerance, insect resistance, pest resistance,disease resistance, and abiotic stress resistance.
 11. The plant, plantpart, or plant cell of claim 7, wherein the single locus that confersherbicide tolerance confers tolerance to benzonitrile herbicides,cyclohexanedione herbicides, imidazolinone herbicides, phenoxyherbicides, sulfonylurea herbicides, triazine herbicides,1-aminocyclopropane-1-carboxylic acid synthase-inhibiting herbicides,4-hydroxyphenylpyruvate dioxygenase-inhibiting herbicides, acetolactatesynthase-inhibiting herbicides, protoporphyrinogen oxidase-inhibitingherbicides, 2,4-dichlorophenoxyacetic acid, bromoxynil, dicamba,glufosinate, glyphosate, nicosulfuron, or quizalofop-p-ethyl.
 12. Amethod of producing a commodity plant product, the method comprisingproducing the commodity plant product from the plant, plant part, orplant cell of claim
 1. 13. The method of claim 12, wherein the commodityplant product is selected from a group consisting of processed hempinflorescence, hemp fiber, hemp oil extract, terpenes, and cannabinoids.14. A method of producing a cannabinoid and/or terpene extract, saidmethod comprising the step of contacting the plant, plant part, or plantcell of claim 1 with a solvent, or exposing said plant, plant part, orplant cell to vaporizing heat, thereby producing a cannabinoid and/orterpene extract.
 15. A method of producing a hemp plant, comprisingplacing the seed of claim 1 in conditions conducive to germination,thereby producing a hemp plant.
 16. A method of producing a hemp plantwith cannabidiol, said method comprising propagating a vegetativecutting from the hemp plant variety designated ‘05.09.24.S1’ whereinsaid hemp plant has all the morphological and physiologicalcharacteristics of ‘05.09.24.S1’ deposited under NCMA No.
 202202006. 17.A method for producing a progeny Cannabis seed, comprising crossing thehemp plant of claim 1, with itself or with another plant, therebyproducing a progeny Cannabis seed.
 18. The method of claim 17, furthercomprising the step of growing the progeny Cannabis seed to produce aprogeny Cannabis plant.
 19. The method of claim 18, further comprisingthe steps of: a) crossing the progeny Cannabis plant from a previousstep with itself or another Cannabis plant to produce a progeny Cannabisplant of a subsequent generation; b) repeating step a) for one or moreadditional generations to produce a Cannabis plant further derived fromthe hemp variety designated ‘05.09.24.S1’.
 20. The method of claim 19,further comprising the step of contacting the Cannabis plant furtherderived from the hemp variety designated ‘05.09.24.S1’ or a plant partderived therefrom with a solvent, or exposing said Cannabis plant orplant part to vaporizing heat, thereby producing a cannabinoid and/orterpene extract.
 21. A method for producing a Cannabis plant derivedfrom a hemp variety designated ‘05.09.24. S1’, said method comprising:propagating a vegetative cutting from a stock Cannabis plant, therebyproducing the Cannabis plant derived from the hemp variety designated‘05.09.24.S1’; wherein the stock Cannabis plant is a product of applyinga plant breeding technique to ‘05.09.24.S1’, wherein seed of the‘05.09.24.S1’ variety has been deposited under NCMA No.
 202202006. 22.The method of claim 21, further comprising the step of contacting theCannabis plant derived from the hemp variety designated ‘05.09.24.S1’ ora plant part derived therefrom with a solvent, or exposing said Cannabisplant or plant part to vaporizing heat, thereby producing a cannabinoidand/or terpene extract.
 23. A method for producing a Cannabis plantderived from a hemp variety designated ‘05.09.24.S1’, said methodcomprising: crossing a stock Cannabis plant with itself or anotherCannabis plant, thereby producing the Cannabis plant derived from thehemp variety designated ‘05.09.24.S1’; wherein the stock Cannabis plantis a product of applying a plant breeding technique to ‘05.09.24.S1’,wherein seed of the ‘05.09.24.S1’ variety has been deposited under NCMANo.
 202202006. 24. The method of claim 23, further comprising the stepof contacting the Cannabis plant derived from the hemp varietydesignated ‘05.09.24.S1’ or a plant part derived therefrom with asolvent, or exposing said Cannabis plant or plant part to vaporizingheat, thereby producing a cannabinoid and/or terpene extract.
 25. Themethod of claim 23, wherein said plant breeding technique is recurrentselection.
 26. The method of claim 23, wherein said plant breedingtechnique is mass selection.
 27. The method of claim 23, wherein saidplant breeding technique is hybridization.
 28. The method of claim 23,wherein said plant breeding technique is open-pollination.
 29. Themethod of claim 21, wherein said plant breeding technique ishybridization.
 30. The method of claim 21, wherein said plant breedingtechnique is recurrent selection.